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Research ArticleSystems Pharmacological Approach to the Effect ofBulsu-san Promoting Parturition
Su Yeon Suh andWon G An
Department of Pharmacology School of Korean Medicine Pusan National University Yangsan Gyeongnam 50612 Republic of Korea
Correspondence should be addressed to Won G An wganpusanackr
Received 28 July 2017 Accepted 25 September 2017 Published 29 October 2017
Academic Editor Gihyun Lee
Copyright copy 2017 Su Yeon Suh and Won G AnThis is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in anymedium provided the originalwork is properly cited
Bulsu-san (BSS) has been commonly used in orientalmedicine for pregnant women in East AsiaThe purpose of this researchwas toelucidate the effect of BSS on ease of parturition using a systems-level in silico analytic approach Research results show that BSS ishighly connected to the parturition related pathways biological processes and organs There were numerous interactions betweenmost compounds of BSS and multiple target genes and this was confirmed using herb-compound-target network target-pathwaynetwork and gene ontology analysis Furthermore the mRNA expression of relevant target genes of BSS was elevated significantlyin related organ tissues such as those of the uterus placenta fetus hypothalamus and pituitary gland This study used a networkanalytical approach to demonstrate that Bulsu-san (BSS) is closely related to the parturition related pathways biological processesand organs It is meaningful that this systems-level network analysis result strengthens the basis of clinical applications of BSS onease of parturition
1 Introduction
The name of Bulsu-san (BSS) originated from its therapeuticeffects that help to promote easy labor as if being touchedby merciful Buddharsquos hand [1] BSS is composed of AngelicaeSinensisRadix (Danggui DG) andCnidium officinaleMakino(Cheongung CG) which is one of the most commonlyused herb pairs in Traditional Medicine of East Asia andthe usual component ratio is 2 3 (CG DG) or 1 1 [2]BSS is widely used in womenrsquos medicine in East Asia itsrecognized therapeutic effects are as follows removal ofimpure blood blood making easy parturition accelerationof labor elimination of dead fetus or placenta amelioration ofpain nourishing blood and promoting blood circulation [3]
What is more recent experimental research on the CG-DG herb pair indicated that they affect the nourishment ofblood [4] activate blood circulation and prevent blood stasis[5] In addition the CG-DG herb pair showed significantinhibitory effects on the proliferation and protein synthesis ofvascular smooth muscle cells [6] It was suggested BSS couldaffect the activities of Akt kinase and eNOS by increasingintracellular Ca2+ and reducing ROS levels [7] and regulatemenstruation and provide relief from pain by enabling the
management of uterine smooth muscle contractions [8]Although BSS has therapeutic effects on various pathologicalsymptoms in pregnant or childbearing aged women thisresearch focused on the molecular mechanisms and impactof BSS on easing parturition and the acceleration of labor
In terms of parturition onset numerous studies havedescribed the complex hormone interactions between estro-gen progesterone oxytocin corticosteroid and prostaglan-din Among these corticotrophin releasing hormone (CRH)is regarded as a trigger that initiates the labor [9]Theplacentareleases substantial amounts of CRH which stimulates thepituitary glands of both mother and fetus to secrete adreno-corticotropin hormone [10] This in turn induces the releaseof estrogen precursor which is converted into estrogen bythe placenta that induces smooth muscle contraction [10]Additionally dilatation of cervical connective tissue andsmooth muscle is induced by the following changes a shiftfrom progesterone to estrogen dominance increased respon-siveness to oxytocin via the upregulation of myometrial oxy-tocin receptor increased prostaglandins synthesis in uterusincreased myometrial gap junction formation decreasednitric oxide activity and increased influx of calcium intomyocyte [11]
HindawiEvidence-Based Complementary and Alternative MedicineVolume 2017 Article ID 7236436 15 pageshttpsdoiorg10115520177236436
2 Evidence-Based Complementary and Alternative Medicine
Figure 1 The workflow the network pharmacological approach of Bulsu-san (BSS) namely active compounds screening target fishingnetwork analysis and relevant organ location mapping was performed in this study
The hypothesis of this study was that BSS may promotethe positive-feedback of hormone loops as well as a seriesof myometrial and cervical changes to ease parturition andsafely accelerate labor A network based in silico approachwas used to identify the effect of BSS on parturition relatedsystems and the aim of this study was to elucidate the effect ofBSS on the parturition by system-level analysisTheworkflowof the network pharmacological study is summarized inFigure 1
2 Material and Methods
21 Identification of Active Compounds Compounds in CGand DG were identified using a phytochemical databasethat is the Traditional Chinese Medicine Systems Phar-macology (TCMSP httpibtshkbueduhkLSPtcmspphp)We applied parameters related to absorption distributionmetabolism and excretion (ADME) namely human drug-likeness (DL) [12] oral bioavailability (OB) [13] and Caco-2 permeability (Caco-2) to screen the Potential active com-pounds in BSS [14]
211 Drug-Likeness Evaluation DL helps filter ldquodrug-likerdquocompounds in oriental herbs as DL represents a qualitativeconcept for valuations based on how ldquodrug-likerdquo a prospec-tive compound is [15] Accordingly a high DL may lead toa greater possibility of therapeutic success and compoundswith a higher DL value are more likely to possess certainbiological properties [16] The calculations of DL in TCMSP
database were based on Tanimoto coefficient formula [17] asfollows
119865 (119860 119861) =119860 times 119861
1198602 + 1198612 minus 119860 times 119861 (1)
where 119860 represents the molecular parameters of herbal com-pounds and119861 is the averagemolecular parameters of all com-pounds in theDrugbank database (httpwwwdrugbankca)[18] In the present study we excluded compounds with a DLof lt008 Other previous researches of herbal formulas set ahigher threshold in the range of 01 to 018However we foundout that most compounds of DG have low DL In detail only36 compounds of 125 in DG show higher or equal DL valuethan 008 For this reason this study sets a lower threshold ofDL than other previous researches to see the most potentialtargets of BSS
212 Oral Bioavailability (OB) Prediction OB is defined asthe ratio of active compoundsrsquo absorption into the systemiccirculation which represents the convergence of the ADMEprocess [13] OB values are dependent on drug dissolution inthe gastrointestinal (GI) tract and hepatic and intestinal first-pass metabolism as well as on intestinal membrane perme-ation whichmakes it a major pharmacokinetic parameter fordrug evaluations [16] In this study the OB threshold was setas ge15
213 Caco-2 Permeability Screening Caco-2 permeability isused to predict the absorption of an orally administered
Evidence-Based Complementary and Alternative Medicine 3
drug [14] Surface absorptivity of the small intestine ismaximized with the presence of villi and microvilli for thisreason most orally administered drug absorption occurs inthe small intestine [19] Moreover the movement of orallyadministered drugs across the intestinal epithelial barrierdetermines the rate and extent of human absorption andultimately affects drug bioavailability [20] In the presentstudy compounds with OB DL and Caco-2 values of greaterthan 15 008 and gtminus04 respectively were regarded asactive compounds and subjected to further analysis
214 Lipinskirsquos Rule (LR) Screening In addition the screen-ing standard used was defined based on Lipinskirsquos rule (LR)which identifies druggable compounds as having molecularweight (MW)ofle500Da (MWle 500) chemical compositionwith le5 hydrogen-bond donors le10 hydrogen-bond accep-tors and an octanol-water partition coefficient AlogP of le5 [21] AlogP can be used to estimate local hydrophobicityto produce molecular hydrophobicity maps and to evaluatehydrophobic interactions in protein-ligand complexes [22]Hdon and Hacc are the number of possible hydrogen-bonddonors and acceptors and the hydrogen-bonding capacity ofa drug solute is recognized as a crucial determinant of perme-ability moreover high hydrogen-bonding potential is oftenrelated to lowpermeability and absorption [23] Eventually inthe present studywe selected active compounds satisfying thefollowing criteria OB ge 15 DL ge 008 Caco-2 ge minus04 MWle 500 H-bond donors le 5 H-bond acceptors le 10 AlogP le 5
22 Target Fishing Aside from filtering active compoundswe also sought to identify the molecular targets of theseactive compounds Compound-target interaction profileswere established based on a systematic prediction of mul-tiple drug-target interactions tool which employs randomforest (RF) and support vector machine (SVM) methodsand integrates chemical genomic and pharmacologicalinformation for drug targeting and discovery on a largescale [24] Compound-target interactions satisfying SVMscore ge 08 and RF score ge 07 were selected for furtherstudy Additionally filtered compound-target interactionprofile mapping was performed using the UniProt database(httpwwwuniprotorg) [25]
23 Gene Ontology (GO) Analysis Biological process (BP)of gene ontology (GO) analysis was employed to determinethe biological properties of target genes [26] GO annotationindicates the possibility of direct statistical analysis on genefunction information In this research GO BP terms with119875 values lt 001 were employed and the data was collectedusing the DAVID 68 Gene Functional Classification Tool(httpdavidabccncifcrfgov)
24 Network Construction and Analysis In order to under-stand the multiscale interactions between the active com-pounds of BSS and targets two types of networks werebuilt (1) the herb-compound-target network (H-C-T net-work) in which nodes represent either compounds tar-get genes or herbs and edges indicate herb-compound-target connections and (2) the target-pathway network (T-P
network) to extract the pathways from KEGG database(httpwwwgenomejpkegg) and the terms highly associ-ated with parturition with 119875 values lt 005 were selected asthe related pathways of targets in this work Related targetswere mapped onto relevant pathways which resulted in theT-P network Both networks were generated in Cytoscape351 an open-source biological network visualization anddata integration software package [27]
25 Target Organ Location Map Tissue-specific patterns ofmRNA expression can indicate important associations withbiological events or gene functions [28] To explore thebeneficial effects of BSS during parturition it is importantthat the tissue mRNA expression profiles of target genesat the organ level be known [29] The target organ loca-tion map was built according to the Dataset GeneAtlasU133A gcrma (httpbiogpsorg) BioGPS database providesexpression data acquired by direct measurements of geneexpression obtained by microarrays analysis [30] First themRNA expression patterns of each target gene in 176 partsof organ tissues were obtained Second average values werecalculated for each gene Third frequency of above averagemRNA expression tissue organs was inspected Forth basedon the result from the third step and parturition mechanismtheory mRNA expression data of relevant organ tissueswere extracted and categorized into 6 groups namely uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smooth muscle and whole blood
3 Results
31 Identification of Active Compounds 314 compounds ofBSSwere identified including 189molecules in CG and 125 inDG (as shown in Supplementary Material Table S1 in Supple-mentary Material available online at httpsdoiorg10115520177236436) and active compounds met the criteria OBge 15 Caco-2 ge minus04 and DL ge 008 as well as thestandards of Lipinskirsquos rule (LR) (as shown in Table 1) Indetail 60 active compounds were initially chosen but 8 com-pounds were present in both herbs namely 3-butylidene-7-hydroxyphthalide adenine BdPh beta-selinene palmiticacid senkyunolide-C senkyunolide-D and senkyunolide-E and 14 had no target protein information and werethus excluded from the list of active compounds whereas27 compounds with lower ADME properties than abovethresholds were included which were reported to be relatedto oxytocin In total 65 active compounds were filtered
Although ligustilide and ferulic acid have a DL of lt008both were included in this study Since ligustilide (C12 DL =007 OB = 5372 Caco-2 = 13) was reported to be the maincompound of DG in uterine contraction [31] and ferulic acid(C42DL=006OB=5497 Caco-2= 053) has been reportedto be useful for the treatment of vascular diseases [6 32] andblood deficiency syndrome [33] in China and to suppressinflammatory responses and tumor progression [34] Someother compounds also have been shown experimentally tohave various biological activities for example crysophanol(C42 DL = 021 OB = 1864 Caco-2 = 062) can be usedto treat menorrhagia and thrombocytopenia [35] Perlolyrine
4 Evidence-Based Complementary and Alternative Medicine
Table 1 65 Potential active compounds of BSS (compound with lowast was present in both herbs)
(C52 DL = 027 OB = 6595 Caco-2 = 088) was confirmedto have a protective effect on injured human umbilical veinendothelial cells [36] and myricanone (C48 DL = 051 OB= 5761 Caco-2 = 067) was found to best inhibit mouse skintumor progression [37]
32 Target Fishing The 65 active compounds interact with185 target proteins as shown in Table 2 in other wordson average each compound on average interacts with 285target proteinsThis result confirms the polypharmacologicalcharacter of oriental medicine and demonstrates the syn-ergistic effects of multiple compounds on multiple targets[38] Different compounds in CG and DG can directly affectcommon targets for example the target protein ldquocalmodulin(CALM1)rdquo interacts with crysophanol fromCG and coniferylferulate from DG at the same time which implies thesynergetic or cumulative effects of herbal medicine
33 GO Analysis 397 biological process terms with 119875 valuesof lt001 were sorted using the functional annotation chart ofthe DAVID 68 Gene Functional Classification Tool basedon 185 filtered target genes and 119875 values were adjustedusing the Benjamini-Hochberg method 30 enriched GO BPterms extracted by 119875 value and gene counts are displayed inFigure 2 It is meaningful that most of the target genes aresignificantly related to the various BP involved in parturitionFor instance 30 extracted GO BP terms include ldquoMAPKsignaling pathwaysrdquo ldquosteroid hormone mediated signalingpathwayrdquo ldquoresponse to glucocorticoidrdquo ldquoresponse to estra-diolrdquo and ldquopositive regulation of ERK1 and ERK2 cascaderdquoldquoMAPK signaling pathwaysrdquo were reported to be activatedin human uterine cervical ripening during parturition [39]ldquoSteroid hormone mediated signaling pathwayrdquo is highly
related to parturition process as estrogen and progesteroneplay important roles in pregnancy and parturition andestrogen induceS the principal stimulatory myometrial con-tractility [40] Also estradiol takes key place in parturitionprocess [41] It was identified that increased ERK activationis observed at the onset of labor and it promotes myometrialcontractility and development of parturition [42 43] To sumup the target genes of BSS are highly associated with thebiological process (BP) of parturition
34 Network Construction and Analysis Network analysis isan efficient tool for visualizing and understanding multipletargeted drug actions and demonstrates drug actions withinthe context of the whole genome [44 45] For a better insightof therapeutic impacts H-C-T and T-P networks were con-structed and displayed in Figures 3 and 4 respectively In theH-C-T network nodes represent herb names compoundsand targets Also in the T-P network circular nodes representtargets and triangle nodes represent pathways Besides nodesize is relative to the degree and edges show interactionsbetween nodes
H-C-T network confirmed that there were 739 interac-tions between 185 targets and 65 active compounds of CGand DG oleic acid (C48 degree = 42) with the highestnumber of interactions with targets followed by succinicacid (C63 degree = 40) and stigmasterol (C62 degree =37) It shows that single molecules target multiple recep-tors [46] Also some compounds from CG and DG werefound to share common targets Likewise prostaglandinGH synthase 2 (PTGS2 degree = 56) displayed the mostaffinitive connections with compounds followed by gamma-aminobutyric acid receptor subunit alpha-1 (GABRA1 degree= 48) prostaglandin GH synthase 1 (PTGS1 degree = 37)
6 Evidence-Based Complementary and Alternative Medicine
Table 2 Related targets of potential compounds in BSS
Evidence-Based Complementary and Alternative Medicine 9
Table 2 Continued
UniProt ID Target name Gene NameP19793 Retinoic acid receptor RXR-alpha RXRAO00767 Acyl-CoA desaturase SCDQ14524 Sodium channel protein type 5 subunit alpha SCN5AP31040 Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial SDHAP16109 P-selectin SELPP05121 Plasminogen activator inhibitor 1 SERPINE1P14410 Sucrase-isomaltase intestinal SIO76082 Solute carrier family 22 member 5 SLC22A5Q9UBX3 Mitochondrial dicarboxylate carrier SLC25A10P11168 Solute carrier family 2 facilitated glucose transporter member 2 SLC2A2P23975 Sodium-dependent noradrenaline transporter SLC6A2Q01959 Sodium-dependent dopamine transporter SLC6A3P31645 Sodium-dependent serotonin transporter SLC6A4P35610 Sterol O-acyltransferase 1 SOAT1P00441 Superoxide dismutase [Cu-Zn] SOD1P08047 Transcription factor Sp1 SP1P12931 Proto-oncogene tyrosine-protein kinase SRC SRCP36956 Sterol regulatory element-binding protein 1 SREBF1Q12772 Sterol regulatory element-binding protein 2 SREBF2Q9P2R7 Succinyl-CoA ligase [ADP-forming] beta-chain mitochondrial SUCLA2Q99973 Telomerase protein component 1 TEP1P01375 Tumor necrosis factor TNFQ16881 Thioredoxin reductase cytoplasmic TXNRD1P55851 Mitochondrial uncoupling protein 2 UCP2P55916 Mitochondrial uncoupling protein 3 UCP3
and muscarinic acetylcholine receptor M1 (CHRM1 degree= 37) Except for C60 (PLA2G1B degree = 1) the rest ofthe 64 active compounds are connected with more thanone target likewise 73 (395) target genes out of 185interactedwithmore than one compoundThis result demon-strates the multicompounds and multitarget properties ofherbal compounds and there was a report that compoundswith multiple targets could have greater therapeutic efficacy[47]
In addition the top 40 pathways were extracted basedon gene counts and 119875 value (lt005) and 119875 value wasadjusted by Benjamini-Hochbergmethod T-P network usingrelevant targets of herbal compounds is demonstrated inFigure 4 There were 485 interactions between the top 40pathways and 135 of 185 target genes ldquoMetabolic pathwaysrdquo(degree = 49) and ldquoneuroactive ligand-receptor interactionpathwayrdquo (degree = 32) had the highest and the secondhighest numbers of connections with the targets followedby ldquocalcium signalingrdquo (degree = 21) ldquocAMP signaling path-wayrdquo (degree = 17) and ldquocGMP PKG signaling pathwayrdquo(degree = 15) These are compelling results that parturitionprocesses are the complex hormone interactions and it iswell known that calcium signals within the myometrium arepivotal for uterine contractions [48] In the same mannersome target genes demonstrated higher degree centralitywith top 40 pathways namely PI3-kinase subunit gamma(PIK3CG degree = 23) cAMP-dependent protein kinase
catalytic subunit alpha (PRKACA degree = 20) proteinkinase C beta type (PRKCB degree = 18) and calmodulin(CALM1 degree = 11) We can confirm the same result in theprevious researches For instance PI3-kinase subunit gammaplays the key role in regulating cAMP calcium cyclingand beta-adrenergic signaling [49] Moreover during thelabor calmodulin-calcium complex activates myosin light-chain kinase which causes the generation of ATPase activityeventually uterine contraction is promoted [50]
H-C-T network explains the multitarget multicom-pounds properties and accumulates effect of herbalmedicines and T-P network shows that target genes of BSSare highly related to the pathway associated with parturi-tion process
35 Target Organ Location Map It is important to confirmthe tissue mRNA expression profiles of the target genes atthe organ level to identify the effects of BSS on parturi-tion Since there was no mRNA expression information inBioGPS of muscarinic acetylcholine receptor M1 (CHRM1)putative beta-glucuronidase-like protein SMA3 (GUSBP1)and retinol-binding protein 2 (RBP2) excluding these 3targets from 185 filtered targets totally 182 genes mRNAexpression profiles were analyzed in this studyTherewere 519interactions between target genes and organ locations Thenetworks of target genes tissuemRNAexpression profiles andcompounds of BSS are shown in Figure 5
10 Evidence-Based Complementary and Alternative Medicine
GO Analysis
688
79
89
899
88910111111
1012
915
1219
1820
2218
2820
34
61 48 49
59 46
56 48 60 55 56 70 70 62 54 47 50 51 63
62 110
62 101
50 64
62 58
117 53
136 233
0 10 20 30 40 50response to fatty acid
circadian rhythmresponse to nutrient
glucose transportcellular response to lipopolysaccharide
steroid hormone mediated signaling pathwayresponse to ethanol
response to hydrogen peroxidepositive regulation of protein kinase B signaling
positive regulation of MAPK cascaderesponse to nicotine
response to coldcholesterol metabolic process
platelet activationpositive regulation of ERK1 and ERK2 cascade
response to lipopolysaccharidelipid metabolic process
response to estradioltranscription initiation from RNA polymerase II promoter
cell-cell signalingresponse to glucocorticoid
positive regulation of transcription DNA-templatedcell proliferation
positive regulation of cell proliferationoxidation-reduction process
positive regulation of transcription from RNA polymerase II promoter
Figure 2GOanalysis 30 enriched biological process (BP) of gene ontology (GO) terms sorted by119875 valuelt 001 and gene counts are displayedThe 119910-axis represents enriched biological process (BP) terms for the target genes and the 119909-axis shows gene counts and minus log 10 (119875 value)
As a result 159 of 182 target genes displayed beyond aver-agemRNAexpression in relevant organ tissues such as uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smoothmuscle andwhole bloodThe rest of23 genes of 182 targets did not display above average mRNAexpression in above organ tissues for example gamma-aminobutyric acid receptor subunit alpha-6 (GABRA6) andcoagulation factor X (F10)
Nevertheless most genes of 159 demonstrated highexpression patterns in several organs of parturition relatedtissues at the same time In detail 60 genes showed mostsignificantmRNA expression in the uterus andor uterus cor-pus group 130 for placenta andor fetus 86 for hypothalamusandor pituitary 82 for smoothmuscle 80 for pituitary and 81for whole blood Besides 30 of 159 genes showed expression
in all of 6 groups For instance muscarinic acetylcholinereceptor M2 (CHRM2) neuronal acetylcholine receptorsubunit 120572-2 (CHRNA2) gamma-aminobutyric acid receptorsubunit alpha-3 (GABRA3) NO synthase inducible (NOS2)cGMP-inhibited 3101584051015840-cyclic phosphodiesterase A (PDE3A)and sodium-dependent dopamine transporter (SLC6A3)recorded beyond average mRNA expression in all six groupsFurthermore 79 of targets were expressed in two or moreorgan tissues which suggests that those organs and targetgenes of BSS are closely correlated
4 Discussion
In this study network pharmacology method with DL OBCaco-2 and LR evaluation multiple drug-target prediction
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
Figure 1 The workflow the network pharmacological approach of Bulsu-san (BSS) namely active compounds screening target fishingnetwork analysis and relevant organ location mapping was performed in this study
The hypothesis of this study was that BSS may promotethe positive-feedback of hormone loops as well as a seriesof myometrial and cervical changes to ease parturition andsafely accelerate labor A network based in silico approachwas used to identify the effect of BSS on parturition relatedsystems and the aim of this study was to elucidate the effect ofBSS on the parturition by system-level analysisTheworkflowof the network pharmacological study is summarized inFigure 1
2 Material and Methods
21 Identification of Active Compounds Compounds in CGand DG were identified using a phytochemical databasethat is the Traditional Chinese Medicine Systems Phar-macology (TCMSP httpibtshkbueduhkLSPtcmspphp)We applied parameters related to absorption distributionmetabolism and excretion (ADME) namely human drug-likeness (DL) [12] oral bioavailability (OB) [13] and Caco-2 permeability (Caco-2) to screen the Potential active com-pounds in BSS [14]
211 Drug-Likeness Evaluation DL helps filter ldquodrug-likerdquocompounds in oriental herbs as DL represents a qualitativeconcept for valuations based on how ldquodrug-likerdquo a prospec-tive compound is [15] Accordingly a high DL may lead toa greater possibility of therapeutic success and compoundswith a higher DL value are more likely to possess certainbiological properties [16] The calculations of DL in TCMSP
database were based on Tanimoto coefficient formula [17] asfollows
119865 (119860 119861) =119860 times 119861
1198602 + 1198612 minus 119860 times 119861 (1)
where 119860 represents the molecular parameters of herbal com-pounds and119861 is the averagemolecular parameters of all com-pounds in theDrugbank database (httpwwwdrugbankca)[18] In the present study we excluded compounds with a DLof lt008 Other previous researches of herbal formulas set ahigher threshold in the range of 01 to 018However we foundout that most compounds of DG have low DL In detail only36 compounds of 125 in DG show higher or equal DL valuethan 008 For this reason this study sets a lower threshold ofDL than other previous researches to see the most potentialtargets of BSS
212 Oral Bioavailability (OB) Prediction OB is defined asthe ratio of active compoundsrsquo absorption into the systemiccirculation which represents the convergence of the ADMEprocess [13] OB values are dependent on drug dissolution inthe gastrointestinal (GI) tract and hepatic and intestinal first-pass metabolism as well as on intestinal membrane perme-ation whichmakes it a major pharmacokinetic parameter fordrug evaluations [16] In this study the OB threshold was setas ge15
213 Caco-2 Permeability Screening Caco-2 permeability isused to predict the absorption of an orally administered
Evidence-Based Complementary and Alternative Medicine 3
drug [14] Surface absorptivity of the small intestine ismaximized with the presence of villi and microvilli for thisreason most orally administered drug absorption occurs inthe small intestine [19] Moreover the movement of orallyadministered drugs across the intestinal epithelial barrierdetermines the rate and extent of human absorption andultimately affects drug bioavailability [20] In the presentstudy compounds with OB DL and Caco-2 values of greaterthan 15 008 and gtminus04 respectively were regarded asactive compounds and subjected to further analysis
214 Lipinskirsquos Rule (LR) Screening In addition the screen-ing standard used was defined based on Lipinskirsquos rule (LR)which identifies druggable compounds as having molecularweight (MW)ofle500Da (MWle 500) chemical compositionwith le5 hydrogen-bond donors le10 hydrogen-bond accep-tors and an octanol-water partition coefficient AlogP of le5 [21] AlogP can be used to estimate local hydrophobicityto produce molecular hydrophobicity maps and to evaluatehydrophobic interactions in protein-ligand complexes [22]Hdon and Hacc are the number of possible hydrogen-bonddonors and acceptors and the hydrogen-bonding capacity ofa drug solute is recognized as a crucial determinant of perme-ability moreover high hydrogen-bonding potential is oftenrelated to lowpermeability and absorption [23] Eventually inthe present studywe selected active compounds satisfying thefollowing criteria OB ge 15 DL ge 008 Caco-2 ge minus04 MWle 500 H-bond donors le 5 H-bond acceptors le 10 AlogP le 5
22 Target Fishing Aside from filtering active compoundswe also sought to identify the molecular targets of theseactive compounds Compound-target interaction profileswere established based on a systematic prediction of mul-tiple drug-target interactions tool which employs randomforest (RF) and support vector machine (SVM) methodsand integrates chemical genomic and pharmacologicalinformation for drug targeting and discovery on a largescale [24] Compound-target interactions satisfying SVMscore ge 08 and RF score ge 07 were selected for furtherstudy Additionally filtered compound-target interactionprofile mapping was performed using the UniProt database(httpwwwuniprotorg) [25]
23 Gene Ontology (GO) Analysis Biological process (BP)of gene ontology (GO) analysis was employed to determinethe biological properties of target genes [26] GO annotationindicates the possibility of direct statistical analysis on genefunction information In this research GO BP terms with119875 values lt 001 were employed and the data was collectedusing the DAVID 68 Gene Functional Classification Tool(httpdavidabccncifcrfgov)
24 Network Construction and Analysis In order to under-stand the multiscale interactions between the active com-pounds of BSS and targets two types of networks werebuilt (1) the herb-compound-target network (H-C-T net-work) in which nodes represent either compounds tar-get genes or herbs and edges indicate herb-compound-target connections and (2) the target-pathway network (T-P
network) to extract the pathways from KEGG database(httpwwwgenomejpkegg) and the terms highly associ-ated with parturition with 119875 values lt 005 were selected asthe related pathways of targets in this work Related targetswere mapped onto relevant pathways which resulted in theT-P network Both networks were generated in Cytoscape351 an open-source biological network visualization anddata integration software package [27]
25 Target Organ Location Map Tissue-specific patterns ofmRNA expression can indicate important associations withbiological events or gene functions [28] To explore thebeneficial effects of BSS during parturition it is importantthat the tissue mRNA expression profiles of target genesat the organ level be known [29] The target organ loca-tion map was built according to the Dataset GeneAtlasU133A gcrma (httpbiogpsorg) BioGPS database providesexpression data acquired by direct measurements of geneexpression obtained by microarrays analysis [30] First themRNA expression patterns of each target gene in 176 partsof organ tissues were obtained Second average values werecalculated for each gene Third frequency of above averagemRNA expression tissue organs was inspected Forth basedon the result from the third step and parturition mechanismtheory mRNA expression data of relevant organ tissueswere extracted and categorized into 6 groups namely uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smooth muscle and whole blood
3 Results
31 Identification of Active Compounds 314 compounds ofBSSwere identified including 189molecules in CG and 125 inDG (as shown in Supplementary Material Table S1 in Supple-mentary Material available online at httpsdoiorg10115520177236436) and active compounds met the criteria OBge 15 Caco-2 ge minus04 and DL ge 008 as well as thestandards of Lipinskirsquos rule (LR) (as shown in Table 1) Indetail 60 active compounds were initially chosen but 8 com-pounds were present in both herbs namely 3-butylidene-7-hydroxyphthalide adenine BdPh beta-selinene palmiticacid senkyunolide-C senkyunolide-D and senkyunolide-E and 14 had no target protein information and werethus excluded from the list of active compounds whereas27 compounds with lower ADME properties than abovethresholds were included which were reported to be relatedto oxytocin In total 65 active compounds were filtered
Although ligustilide and ferulic acid have a DL of lt008both were included in this study Since ligustilide (C12 DL =007 OB = 5372 Caco-2 = 13) was reported to be the maincompound of DG in uterine contraction [31] and ferulic acid(C42DL=006OB=5497 Caco-2= 053) has been reportedto be useful for the treatment of vascular diseases [6 32] andblood deficiency syndrome [33] in China and to suppressinflammatory responses and tumor progression [34] Someother compounds also have been shown experimentally tohave various biological activities for example crysophanol(C42 DL = 021 OB = 1864 Caco-2 = 062) can be usedto treat menorrhagia and thrombocytopenia [35] Perlolyrine
4 Evidence-Based Complementary and Alternative Medicine
Table 1 65 Potential active compounds of BSS (compound with lowast was present in both herbs)
(C52 DL = 027 OB = 6595 Caco-2 = 088) was confirmedto have a protective effect on injured human umbilical veinendothelial cells [36] and myricanone (C48 DL = 051 OB= 5761 Caco-2 = 067) was found to best inhibit mouse skintumor progression [37]
32 Target Fishing The 65 active compounds interact with185 target proteins as shown in Table 2 in other wordson average each compound on average interacts with 285target proteinsThis result confirms the polypharmacologicalcharacter of oriental medicine and demonstrates the syn-ergistic effects of multiple compounds on multiple targets[38] Different compounds in CG and DG can directly affectcommon targets for example the target protein ldquocalmodulin(CALM1)rdquo interacts with crysophanol fromCG and coniferylferulate from DG at the same time which implies thesynergetic or cumulative effects of herbal medicine
33 GO Analysis 397 biological process terms with 119875 valuesof lt001 were sorted using the functional annotation chart ofthe DAVID 68 Gene Functional Classification Tool basedon 185 filtered target genes and 119875 values were adjustedusing the Benjamini-Hochberg method 30 enriched GO BPterms extracted by 119875 value and gene counts are displayed inFigure 2 It is meaningful that most of the target genes aresignificantly related to the various BP involved in parturitionFor instance 30 extracted GO BP terms include ldquoMAPKsignaling pathwaysrdquo ldquosteroid hormone mediated signalingpathwayrdquo ldquoresponse to glucocorticoidrdquo ldquoresponse to estra-diolrdquo and ldquopositive regulation of ERK1 and ERK2 cascaderdquoldquoMAPK signaling pathwaysrdquo were reported to be activatedin human uterine cervical ripening during parturition [39]ldquoSteroid hormone mediated signaling pathwayrdquo is highly
related to parturition process as estrogen and progesteroneplay important roles in pregnancy and parturition andestrogen induceS the principal stimulatory myometrial con-tractility [40] Also estradiol takes key place in parturitionprocess [41] It was identified that increased ERK activationis observed at the onset of labor and it promotes myometrialcontractility and development of parturition [42 43] To sumup the target genes of BSS are highly associated with thebiological process (BP) of parturition
34 Network Construction and Analysis Network analysis isan efficient tool for visualizing and understanding multipletargeted drug actions and demonstrates drug actions withinthe context of the whole genome [44 45] For a better insightof therapeutic impacts H-C-T and T-P networks were con-structed and displayed in Figures 3 and 4 respectively In theH-C-T network nodes represent herb names compoundsand targets Also in the T-P network circular nodes representtargets and triangle nodes represent pathways Besides nodesize is relative to the degree and edges show interactionsbetween nodes
H-C-T network confirmed that there were 739 interac-tions between 185 targets and 65 active compounds of CGand DG oleic acid (C48 degree = 42) with the highestnumber of interactions with targets followed by succinicacid (C63 degree = 40) and stigmasterol (C62 degree =37) It shows that single molecules target multiple recep-tors [46] Also some compounds from CG and DG werefound to share common targets Likewise prostaglandinGH synthase 2 (PTGS2 degree = 56) displayed the mostaffinitive connections with compounds followed by gamma-aminobutyric acid receptor subunit alpha-1 (GABRA1 degree= 48) prostaglandin GH synthase 1 (PTGS1 degree = 37)
6 Evidence-Based Complementary and Alternative Medicine
Table 2 Related targets of potential compounds in BSS
Evidence-Based Complementary and Alternative Medicine 9
Table 2 Continued
UniProt ID Target name Gene NameP19793 Retinoic acid receptor RXR-alpha RXRAO00767 Acyl-CoA desaturase SCDQ14524 Sodium channel protein type 5 subunit alpha SCN5AP31040 Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial SDHAP16109 P-selectin SELPP05121 Plasminogen activator inhibitor 1 SERPINE1P14410 Sucrase-isomaltase intestinal SIO76082 Solute carrier family 22 member 5 SLC22A5Q9UBX3 Mitochondrial dicarboxylate carrier SLC25A10P11168 Solute carrier family 2 facilitated glucose transporter member 2 SLC2A2P23975 Sodium-dependent noradrenaline transporter SLC6A2Q01959 Sodium-dependent dopamine transporter SLC6A3P31645 Sodium-dependent serotonin transporter SLC6A4P35610 Sterol O-acyltransferase 1 SOAT1P00441 Superoxide dismutase [Cu-Zn] SOD1P08047 Transcription factor Sp1 SP1P12931 Proto-oncogene tyrosine-protein kinase SRC SRCP36956 Sterol regulatory element-binding protein 1 SREBF1Q12772 Sterol regulatory element-binding protein 2 SREBF2Q9P2R7 Succinyl-CoA ligase [ADP-forming] beta-chain mitochondrial SUCLA2Q99973 Telomerase protein component 1 TEP1P01375 Tumor necrosis factor TNFQ16881 Thioredoxin reductase cytoplasmic TXNRD1P55851 Mitochondrial uncoupling protein 2 UCP2P55916 Mitochondrial uncoupling protein 3 UCP3
and muscarinic acetylcholine receptor M1 (CHRM1 degree= 37) Except for C60 (PLA2G1B degree = 1) the rest ofthe 64 active compounds are connected with more thanone target likewise 73 (395) target genes out of 185interactedwithmore than one compoundThis result demon-strates the multicompounds and multitarget properties ofherbal compounds and there was a report that compoundswith multiple targets could have greater therapeutic efficacy[47]
In addition the top 40 pathways were extracted basedon gene counts and 119875 value (lt005) and 119875 value wasadjusted by Benjamini-Hochbergmethod T-P network usingrelevant targets of herbal compounds is demonstrated inFigure 4 There were 485 interactions between the top 40pathways and 135 of 185 target genes ldquoMetabolic pathwaysrdquo(degree = 49) and ldquoneuroactive ligand-receptor interactionpathwayrdquo (degree = 32) had the highest and the secondhighest numbers of connections with the targets followedby ldquocalcium signalingrdquo (degree = 21) ldquocAMP signaling path-wayrdquo (degree = 17) and ldquocGMP PKG signaling pathwayrdquo(degree = 15) These are compelling results that parturitionprocesses are the complex hormone interactions and it iswell known that calcium signals within the myometrium arepivotal for uterine contractions [48] In the same mannersome target genes demonstrated higher degree centralitywith top 40 pathways namely PI3-kinase subunit gamma(PIK3CG degree = 23) cAMP-dependent protein kinase
catalytic subunit alpha (PRKACA degree = 20) proteinkinase C beta type (PRKCB degree = 18) and calmodulin(CALM1 degree = 11) We can confirm the same result in theprevious researches For instance PI3-kinase subunit gammaplays the key role in regulating cAMP calcium cyclingand beta-adrenergic signaling [49] Moreover during thelabor calmodulin-calcium complex activates myosin light-chain kinase which causes the generation of ATPase activityeventually uterine contraction is promoted [50]
H-C-T network explains the multitarget multicom-pounds properties and accumulates effect of herbalmedicines and T-P network shows that target genes of BSSare highly related to the pathway associated with parturi-tion process
35 Target Organ Location Map It is important to confirmthe tissue mRNA expression profiles of the target genes atthe organ level to identify the effects of BSS on parturi-tion Since there was no mRNA expression information inBioGPS of muscarinic acetylcholine receptor M1 (CHRM1)putative beta-glucuronidase-like protein SMA3 (GUSBP1)and retinol-binding protein 2 (RBP2) excluding these 3targets from 185 filtered targets totally 182 genes mRNAexpression profiles were analyzed in this studyTherewere 519interactions between target genes and organ locations Thenetworks of target genes tissuemRNAexpression profiles andcompounds of BSS are shown in Figure 5
10 Evidence-Based Complementary and Alternative Medicine
GO Analysis
688
79
89
899
88910111111
1012
915
1219
1820
2218
2820
34
61 48 49
59 46
56 48 60 55 56 70 70 62 54 47 50 51 63
62 110
62 101
50 64
62 58
117 53
136 233
0 10 20 30 40 50response to fatty acid
circadian rhythmresponse to nutrient
glucose transportcellular response to lipopolysaccharide
steroid hormone mediated signaling pathwayresponse to ethanol
response to hydrogen peroxidepositive regulation of protein kinase B signaling
positive regulation of MAPK cascaderesponse to nicotine
response to coldcholesterol metabolic process
platelet activationpositive regulation of ERK1 and ERK2 cascade
response to lipopolysaccharidelipid metabolic process
response to estradioltranscription initiation from RNA polymerase II promoter
cell-cell signalingresponse to glucocorticoid
positive regulation of transcription DNA-templatedcell proliferation
positive regulation of cell proliferationoxidation-reduction process
positive regulation of transcription from RNA polymerase II promoter
Figure 2GOanalysis 30 enriched biological process (BP) of gene ontology (GO) terms sorted by119875 valuelt 001 and gene counts are displayedThe 119910-axis represents enriched biological process (BP) terms for the target genes and the 119909-axis shows gene counts and minus log 10 (119875 value)
As a result 159 of 182 target genes displayed beyond aver-agemRNAexpression in relevant organ tissues such as uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smoothmuscle andwhole bloodThe rest of23 genes of 182 targets did not display above average mRNAexpression in above organ tissues for example gamma-aminobutyric acid receptor subunit alpha-6 (GABRA6) andcoagulation factor X (F10)
Nevertheless most genes of 159 demonstrated highexpression patterns in several organs of parturition relatedtissues at the same time In detail 60 genes showed mostsignificantmRNA expression in the uterus andor uterus cor-pus group 130 for placenta andor fetus 86 for hypothalamusandor pituitary 82 for smoothmuscle 80 for pituitary and 81for whole blood Besides 30 of 159 genes showed expression
in all of 6 groups For instance muscarinic acetylcholinereceptor M2 (CHRM2) neuronal acetylcholine receptorsubunit 120572-2 (CHRNA2) gamma-aminobutyric acid receptorsubunit alpha-3 (GABRA3) NO synthase inducible (NOS2)cGMP-inhibited 3101584051015840-cyclic phosphodiesterase A (PDE3A)and sodium-dependent dopamine transporter (SLC6A3)recorded beyond average mRNA expression in all six groupsFurthermore 79 of targets were expressed in two or moreorgan tissues which suggests that those organs and targetgenes of BSS are closely correlated
4 Discussion
In this study network pharmacology method with DL OBCaco-2 and LR evaluation multiple drug-target prediction
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
Evidence-Based Complementary and Alternative Medicine 3
drug [14] Surface absorptivity of the small intestine ismaximized with the presence of villi and microvilli for thisreason most orally administered drug absorption occurs inthe small intestine [19] Moreover the movement of orallyadministered drugs across the intestinal epithelial barrierdetermines the rate and extent of human absorption andultimately affects drug bioavailability [20] In the presentstudy compounds with OB DL and Caco-2 values of greaterthan 15 008 and gtminus04 respectively were regarded asactive compounds and subjected to further analysis
214 Lipinskirsquos Rule (LR) Screening In addition the screen-ing standard used was defined based on Lipinskirsquos rule (LR)which identifies druggable compounds as having molecularweight (MW)ofle500Da (MWle 500) chemical compositionwith le5 hydrogen-bond donors le10 hydrogen-bond accep-tors and an octanol-water partition coefficient AlogP of le5 [21] AlogP can be used to estimate local hydrophobicityto produce molecular hydrophobicity maps and to evaluatehydrophobic interactions in protein-ligand complexes [22]Hdon and Hacc are the number of possible hydrogen-bonddonors and acceptors and the hydrogen-bonding capacity ofa drug solute is recognized as a crucial determinant of perme-ability moreover high hydrogen-bonding potential is oftenrelated to lowpermeability and absorption [23] Eventually inthe present studywe selected active compounds satisfying thefollowing criteria OB ge 15 DL ge 008 Caco-2 ge minus04 MWle 500 H-bond donors le 5 H-bond acceptors le 10 AlogP le 5
22 Target Fishing Aside from filtering active compoundswe also sought to identify the molecular targets of theseactive compounds Compound-target interaction profileswere established based on a systematic prediction of mul-tiple drug-target interactions tool which employs randomforest (RF) and support vector machine (SVM) methodsand integrates chemical genomic and pharmacologicalinformation for drug targeting and discovery on a largescale [24] Compound-target interactions satisfying SVMscore ge 08 and RF score ge 07 were selected for furtherstudy Additionally filtered compound-target interactionprofile mapping was performed using the UniProt database(httpwwwuniprotorg) [25]
23 Gene Ontology (GO) Analysis Biological process (BP)of gene ontology (GO) analysis was employed to determinethe biological properties of target genes [26] GO annotationindicates the possibility of direct statistical analysis on genefunction information In this research GO BP terms with119875 values lt 001 were employed and the data was collectedusing the DAVID 68 Gene Functional Classification Tool(httpdavidabccncifcrfgov)
24 Network Construction and Analysis In order to under-stand the multiscale interactions between the active com-pounds of BSS and targets two types of networks werebuilt (1) the herb-compound-target network (H-C-T net-work) in which nodes represent either compounds tar-get genes or herbs and edges indicate herb-compound-target connections and (2) the target-pathway network (T-P
network) to extract the pathways from KEGG database(httpwwwgenomejpkegg) and the terms highly associ-ated with parturition with 119875 values lt 005 were selected asthe related pathways of targets in this work Related targetswere mapped onto relevant pathways which resulted in theT-P network Both networks were generated in Cytoscape351 an open-source biological network visualization anddata integration software package [27]
25 Target Organ Location Map Tissue-specific patterns ofmRNA expression can indicate important associations withbiological events or gene functions [28] To explore thebeneficial effects of BSS during parturition it is importantthat the tissue mRNA expression profiles of target genesat the organ level be known [29] The target organ loca-tion map was built according to the Dataset GeneAtlasU133A gcrma (httpbiogpsorg) BioGPS database providesexpression data acquired by direct measurements of geneexpression obtained by microarrays analysis [30] First themRNA expression patterns of each target gene in 176 partsof organ tissues were obtained Second average values werecalculated for each gene Third frequency of above averagemRNA expression tissue organs was inspected Forth basedon the result from the third step and parturition mechanismtheory mRNA expression data of relevant organ tissueswere extracted and categorized into 6 groups namely uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smooth muscle and whole blood
3 Results
31 Identification of Active Compounds 314 compounds ofBSSwere identified including 189molecules in CG and 125 inDG (as shown in Supplementary Material Table S1 in Supple-mentary Material available online at httpsdoiorg10115520177236436) and active compounds met the criteria OBge 15 Caco-2 ge minus04 and DL ge 008 as well as thestandards of Lipinskirsquos rule (LR) (as shown in Table 1) Indetail 60 active compounds were initially chosen but 8 com-pounds were present in both herbs namely 3-butylidene-7-hydroxyphthalide adenine BdPh beta-selinene palmiticacid senkyunolide-C senkyunolide-D and senkyunolide-E and 14 had no target protein information and werethus excluded from the list of active compounds whereas27 compounds with lower ADME properties than abovethresholds were included which were reported to be relatedto oxytocin In total 65 active compounds were filtered
Although ligustilide and ferulic acid have a DL of lt008both were included in this study Since ligustilide (C12 DL =007 OB = 5372 Caco-2 = 13) was reported to be the maincompound of DG in uterine contraction [31] and ferulic acid(C42DL=006OB=5497 Caco-2= 053) has been reportedto be useful for the treatment of vascular diseases [6 32] andblood deficiency syndrome [33] in China and to suppressinflammatory responses and tumor progression [34] Someother compounds also have been shown experimentally tohave various biological activities for example crysophanol(C42 DL = 021 OB = 1864 Caco-2 = 062) can be usedto treat menorrhagia and thrombocytopenia [35] Perlolyrine
4 Evidence-Based Complementary and Alternative Medicine
Table 1 65 Potential active compounds of BSS (compound with lowast was present in both herbs)
(C52 DL = 027 OB = 6595 Caco-2 = 088) was confirmedto have a protective effect on injured human umbilical veinendothelial cells [36] and myricanone (C48 DL = 051 OB= 5761 Caco-2 = 067) was found to best inhibit mouse skintumor progression [37]
32 Target Fishing The 65 active compounds interact with185 target proteins as shown in Table 2 in other wordson average each compound on average interacts with 285target proteinsThis result confirms the polypharmacologicalcharacter of oriental medicine and demonstrates the syn-ergistic effects of multiple compounds on multiple targets[38] Different compounds in CG and DG can directly affectcommon targets for example the target protein ldquocalmodulin(CALM1)rdquo interacts with crysophanol fromCG and coniferylferulate from DG at the same time which implies thesynergetic or cumulative effects of herbal medicine
33 GO Analysis 397 biological process terms with 119875 valuesof lt001 were sorted using the functional annotation chart ofthe DAVID 68 Gene Functional Classification Tool basedon 185 filtered target genes and 119875 values were adjustedusing the Benjamini-Hochberg method 30 enriched GO BPterms extracted by 119875 value and gene counts are displayed inFigure 2 It is meaningful that most of the target genes aresignificantly related to the various BP involved in parturitionFor instance 30 extracted GO BP terms include ldquoMAPKsignaling pathwaysrdquo ldquosteroid hormone mediated signalingpathwayrdquo ldquoresponse to glucocorticoidrdquo ldquoresponse to estra-diolrdquo and ldquopositive regulation of ERK1 and ERK2 cascaderdquoldquoMAPK signaling pathwaysrdquo were reported to be activatedin human uterine cervical ripening during parturition [39]ldquoSteroid hormone mediated signaling pathwayrdquo is highly
related to parturition process as estrogen and progesteroneplay important roles in pregnancy and parturition andestrogen induceS the principal stimulatory myometrial con-tractility [40] Also estradiol takes key place in parturitionprocess [41] It was identified that increased ERK activationis observed at the onset of labor and it promotes myometrialcontractility and development of parturition [42 43] To sumup the target genes of BSS are highly associated with thebiological process (BP) of parturition
34 Network Construction and Analysis Network analysis isan efficient tool for visualizing and understanding multipletargeted drug actions and demonstrates drug actions withinthe context of the whole genome [44 45] For a better insightof therapeutic impacts H-C-T and T-P networks were con-structed and displayed in Figures 3 and 4 respectively In theH-C-T network nodes represent herb names compoundsand targets Also in the T-P network circular nodes representtargets and triangle nodes represent pathways Besides nodesize is relative to the degree and edges show interactionsbetween nodes
H-C-T network confirmed that there were 739 interac-tions between 185 targets and 65 active compounds of CGand DG oleic acid (C48 degree = 42) with the highestnumber of interactions with targets followed by succinicacid (C63 degree = 40) and stigmasterol (C62 degree =37) It shows that single molecules target multiple recep-tors [46] Also some compounds from CG and DG werefound to share common targets Likewise prostaglandinGH synthase 2 (PTGS2 degree = 56) displayed the mostaffinitive connections with compounds followed by gamma-aminobutyric acid receptor subunit alpha-1 (GABRA1 degree= 48) prostaglandin GH synthase 1 (PTGS1 degree = 37)
6 Evidence-Based Complementary and Alternative Medicine
Table 2 Related targets of potential compounds in BSS
Evidence-Based Complementary and Alternative Medicine 9
Table 2 Continued
UniProt ID Target name Gene NameP19793 Retinoic acid receptor RXR-alpha RXRAO00767 Acyl-CoA desaturase SCDQ14524 Sodium channel protein type 5 subunit alpha SCN5AP31040 Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial SDHAP16109 P-selectin SELPP05121 Plasminogen activator inhibitor 1 SERPINE1P14410 Sucrase-isomaltase intestinal SIO76082 Solute carrier family 22 member 5 SLC22A5Q9UBX3 Mitochondrial dicarboxylate carrier SLC25A10P11168 Solute carrier family 2 facilitated glucose transporter member 2 SLC2A2P23975 Sodium-dependent noradrenaline transporter SLC6A2Q01959 Sodium-dependent dopamine transporter SLC6A3P31645 Sodium-dependent serotonin transporter SLC6A4P35610 Sterol O-acyltransferase 1 SOAT1P00441 Superoxide dismutase [Cu-Zn] SOD1P08047 Transcription factor Sp1 SP1P12931 Proto-oncogene tyrosine-protein kinase SRC SRCP36956 Sterol regulatory element-binding protein 1 SREBF1Q12772 Sterol regulatory element-binding protein 2 SREBF2Q9P2R7 Succinyl-CoA ligase [ADP-forming] beta-chain mitochondrial SUCLA2Q99973 Telomerase protein component 1 TEP1P01375 Tumor necrosis factor TNFQ16881 Thioredoxin reductase cytoplasmic TXNRD1P55851 Mitochondrial uncoupling protein 2 UCP2P55916 Mitochondrial uncoupling protein 3 UCP3
and muscarinic acetylcholine receptor M1 (CHRM1 degree= 37) Except for C60 (PLA2G1B degree = 1) the rest ofthe 64 active compounds are connected with more thanone target likewise 73 (395) target genes out of 185interactedwithmore than one compoundThis result demon-strates the multicompounds and multitarget properties ofherbal compounds and there was a report that compoundswith multiple targets could have greater therapeutic efficacy[47]
In addition the top 40 pathways were extracted basedon gene counts and 119875 value (lt005) and 119875 value wasadjusted by Benjamini-Hochbergmethod T-P network usingrelevant targets of herbal compounds is demonstrated inFigure 4 There were 485 interactions between the top 40pathways and 135 of 185 target genes ldquoMetabolic pathwaysrdquo(degree = 49) and ldquoneuroactive ligand-receptor interactionpathwayrdquo (degree = 32) had the highest and the secondhighest numbers of connections with the targets followedby ldquocalcium signalingrdquo (degree = 21) ldquocAMP signaling path-wayrdquo (degree = 17) and ldquocGMP PKG signaling pathwayrdquo(degree = 15) These are compelling results that parturitionprocesses are the complex hormone interactions and it iswell known that calcium signals within the myometrium arepivotal for uterine contractions [48] In the same mannersome target genes demonstrated higher degree centralitywith top 40 pathways namely PI3-kinase subunit gamma(PIK3CG degree = 23) cAMP-dependent protein kinase
catalytic subunit alpha (PRKACA degree = 20) proteinkinase C beta type (PRKCB degree = 18) and calmodulin(CALM1 degree = 11) We can confirm the same result in theprevious researches For instance PI3-kinase subunit gammaplays the key role in regulating cAMP calcium cyclingand beta-adrenergic signaling [49] Moreover during thelabor calmodulin-calcium complex activates myosin light-chain kinase which causes the generation of ATPase activityeventually uterine contraction is promoted [50]
H-C-T network explains the multitarget multicom-pounds properties and accumulates effect of herbalmedicines and T-P network shows that target genes of BSSare highly related to the pathway associated with parturi-tion process
35 Target Organ Location Map It is important to confirmthe tissue mRNA expression profiles of the target genes atthe organ level to identify the effects of BSS on parturi-tion Since there was no mRNA expression information inBioGPS of muscarinic acetylcholine receptor M1 (CHRM1)putative beta-glucuronidase-like protein SMA3 (GUSBP1)and retinol-binding protein 2 (RBP2) excluding these 3targets from 185 filtered targets totally 182 genes mRNAexpression profiles were analyzed in this studyTherewere 519interactions between target genes and organ locations Thenetworks of target genes tissuemRNAexpression profiles andcompounds of BSS are shown in Figure 5
10 Evidence-Based Complementary and Alternative Medicine
GO Analysis
688
79
89
899
88910111111
1012
915
1219
1820
2218
2820
34
61 48 49
59 46
56 48 60 55 56 70 70 62 54 47 50 51 63
62 110
62 101
50 64
62 58
117 53
136 233
0 10 20 30 40 50response to fatty acid
circadian rhythmresponse to nutrient
glucose transportcellular response to lipopolysaccharide
steroid hormone mediated signaling pathwayresponse to ethanol
response to hydrogen peroxidepositive regulation of protein kinase B signaling
positive regulation of MAPK cascaderesponse to nicotine
response to coldcholesterol metabolic process
platelet activationpositive regulation of ERK1 and ERK2 cascade
response to lipopolysaccharidelipid metabolic process
response to estradioltranscription initiation from RNA polymerase II promoter
cell-cell signalingresponse to glucocorticoid
positive regulation of transcription DNA-templatedcell proliferation
positive regulation of cell proliferationoxidation-reduction process
positive regulation of transcription from RNA polymerase II promoter
Figure 2GOanalysis 30 enriched biological process (BP) of gene ontology (GO) terms sorted by119875 valuelt 001 and gene counts are displayedThe 119910-axis represents enriched biological process (BP) terms for the target genes and the 119909-axis shows gene counts and minus log 10 (119875 value)
As a result 159 of 182 target genes displayed beyond aver-agemRNAexpression in relevant organ tissues such as uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smoothmuscle andwhole bloodThe rest of23 genes of 182 targets did not display above average mRNAexpression in above organ tissues for example gamma-aminobutyric acid receptor subunit alpha-6 (GABRA6) andcoagulation factor X (F10)
Nevertheless most genes of 159 demonstrated highexpression patterns in several organs of parturition relatedtissues at the same time In detail 60 genes showed mostsignificantmRNA expression in the uterus andor uterus cor-pus group 130 for placenta andor fetus 86 for hypothalamusandor pituitary 82 for smoothmuscle 80 for pituitary and 81for whole blood Besides 30 of 159 genes showed expression
in all of 6 groups For instance muscarinic acetylcholinereceptor M2 (CHRM2) neuronal acetylcholine receptorsubunit 120572-2 (CHRNA2) gamma-aminobutyric acid receptorsubunit alpha-3 (GABRA3) NO synthase inducible (NOS2)cGMP-inhibited 3101584051015840-cyclic phosphodiesterase A (PDE3A)and sodium-dependent dopamine transporter (SLC6A3)recorded beyond average mRNA expression in all six groupsFurthermore 79 of targets were expressed in two or moreorgan tissues which suggests that those organs and targetgenes of BSS are closely correlated
4 Discussion
In this study network pharmacology method with DL OBCaco-2 and LR evaluation multiple drug-target prediction
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
(C52 DL = 027 OB = 6595 Caco-2 = 088) was confirmedto have a protective effect on injured human umbilical veinendothelial cells [36] and myricanone (C48 DL = 051 OB= 5761 Caco-2 = 067) was found to best inhibit mouse skintumor progression [37]
32 Target Fishing The 65 active compounds interact with185 target proteins as shown in Table 2 in other wordson average each compound on average interacts with 285target proteinsThis result confirms the polypharmacologicalcharacter of oriental medicine and demonstrates the syn-ergistic effects of multiple compounds on multiple targets[38] Different compounds in CG and DG can directly affectcommon targets for example the target protein ldquocalmodulin(CALM1)rdquo interacts with crysophanol fromCG and coniferylferulate from DG at the same time which implies thesynergetic or cumulative effects of herbal medicine
33 GO Analysis 397 biological process terms with 119875 valuesof lt001 were sorted using the functional annotation chart ofthe DAVID 68 Gene Functional Classification Tool basedon 185 filtered target genes and 119875 values were adjustedusing the Benjamini-Hochberg method 30 enriched GO BPterms extracted by 119875 value and gene counts are displayed inFigure 2 It is meaningful that most of the target genes aresignificantly related to the various BP involved in parturitionFor instance 30 extracted GO BP terms include ldquoMAPKsignaling pathwaysrdquo ldquosteroid hormone mediated signalingpathwayrdquo ldquoresponse to glucocorticoidrdquo ldquoresponse to estra-diolrdquo and ldquopositive regulation of ERK1 and ERK2 cascaderdquoldquoMAPK signaling pathwaysrdquo were reported to be activatedin human uterine cervical ripening during parturition [39]ldquoSteroid hormone mediated signaling pathwayrdquo is highly
related to parturition process as estrogen and progesteroneplay important roles in pregnancy and parturition andestrogen induceS the principal stimulatory myometrial con-tractility [40] Also estradiol takes key place in parturitionprocess [41] It was identified that increased ERK activationis observed at the onset of labor and it promotes myometrialcontractility and development of parturition [42 43] To sumup the target genes of BSS are highly associated with thebiological process (BP) of parturition
34 Network Construction and Analysis Network analysis isan efficient tool for visualizing and understanding multipletargeted drug actions and demonstrates drug actions withinthe context of the whole genome [44 45] For a better insightof therapeutic impacts H-C-T and T-P networks were con-structed and displayed in Figures 3 and 4 respectively In theH-C-T network nodes represent herb names compoundsand targets Also in the T-P network circular nodes representtargets and triangle nodes represent pathways Besides nodesize is relative to the degree and edges show interactionsbetween nodes
H-C-T network confirmed that there were 739 interac-tions between 185 targets and 65 active compounds of CGand DG oleic acid (C48 degree = 42) with the highestnumber of interactions with targets followed by succinicacid (C63 degree = 40) and stigmasterol (C62 degree =37) It shows that single molecules target multiple recep-tors [46] Also some compounds from CG and DG werefound to share common targets Likewise prostaglandinGH synthase 2 (PTGS2 degree = 56) displayed the mostaffinitive connections with compounds followed by gamma-aminobutyric acid receptor subunit alpha-1 (GABRA1 degree= 48) prostaglandin GH synthase 1 (PTGS1 degree = 37)
6 Evidence-Based Complementary and Alternative Medicine
Table 2 Related targets of potential compounds in BSS
Evidence-Based Complementary and Alternative Medicine 9
Table 2 Continued
UniProt ID Target name Gene NameP19793 Retinoic acid receptor RXR-alpha RXRAO00767 Acyl-CoA desaturase SCDQ14524 Sodium channel protein type 5 subunit alpha SCN5AP31040 Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial SDHAP16109 P-selectin SELPP05121 Plasminogen activator inhibitor 1 SERPINE1P14410 Sucrase-isomaltase intestinal SIO76082 Solute carrier family 22 member 5 SLC22A5Q9UBX3 Mitochondrial dicarboxylate carrier SLC25A10P11168 Solute carrier family 2 facilitated glucose transporter member 2 SLC2A2P23975 Sodium-dependent noradrenaline transporter SLC6A2Q01959 Sodium-dependent dopamine transporter SLC6A3P31645 Sodium-dependent serotonin transporter SLC6A4P35610 Sterol O-acyltransferase 1 SOAT1P00441 Superoxide dismutase [Cu-Zn] SOD1P08047 Transcription factor Sp1 SP1P12931 Proto-oncogene tyrosine-protein kinase SRC SRCP36956 Sterol regulatory element-binding protein 1 SREBF1Q12772 Sterol regulatory element-binding protein 2 SREBF2Q9P2R7 Succinyl-CoA ligase [ADP-forming] beta-chain mitochondrial SUCLA2Q99973 Telomerase protein component 1 TEP1P01375 Tumor necrosis factor TNFQ16881 Thioredoxin reductase cytoplasmic TXNRD1P55851 Mitochondrial uncoupling protein 2 UCP2P55916 Mitochondrial uncoupling protein 3 UCP3
and muscarinic acetylcholine receptor M1 (CHRM1 degree= 37) Except for C60 (PLA2G1B degree = 1) the rest ofthe 64 active compounds are connected with more thanone target likewise 73 (395) target genes out of 185interactedwithmore than one compoundThis result demon-strates the multicompounds and multitarget properties ofherbal compounds and there was a report that compoundswith multiple targets could have greater therapeutic efficacy[47]
In addition the top 40 pathways were extracted basedon gene counts and 119875 value (lt005) and 119875 value wasadjusted by Benjamini-Hochbergmethod T-P network usingrelevant targets of herbal compounds is demonstrated inFigure 4 There were 485 interactions between the top 40pathways and 135 of 185 target genes ldquoMetabolic pathwaysrdquo(degree = 49) and ldquoneuroactive ligand-receptor interactionpathwayrdquo (degree = 32) had the highest and the secondhighest numbers of connections with the targets followedby ldquocalcium signalingrdquo (degree = 21) ldquocAMP signaling path-wayrdquo (degree = 17) and ldquocGMP PKG signaling pathwayrdquo(degree = 15) These are compelling results that parturitionprocesses are the complex hormone interactions and it iswell known that calcium signals within the myometrium arepivotal for uterine contractions [48] In the same mannersome target genes demonstrated higher degree centralitywith top 40 pathways namely PI3-kinase subunit gamma(PIK3CG degree = 23) cAMP-dependent protein kinase
catalytic subunit alpha (PRKACA degree = 20) proteinkinase C beta type (PRKCB degree = 18) and calmodulin(CALM1 degree = 11) We can confirm the same result in theprevious researches For instance PI3-kinase subunit gammaplays the key role in regulating cAMP calcium cyclingand beta-adrenergic signaling [49] Moreover during thelabor calmodulin-calcium complex activates myosin light-chain kinase which causes the generation of ATPase activityeventually uterine contraction is promoted [50]
H-C-T network explains the multitarget multicom-pounds properties and accumulates effect of herbalmedicines and T-P network shows that target genes of BSSare highly related to the pathway associated with parturi-tion process
35 Target Organ Location Map It is important to confirmthe tissue mRNA expression profiles of the target genes atthe organ level to identify the effects of BSS on parturi-tion Since there was no mRNA expression information inBioGPS of muscarinic acetylcholine receptor M1 (CHRM1)putative beta-glucuronidase-like protein SMA3 (GUSBP1)and retinol-binding protein 2 (RBP2) excluding these 3targets from 185 filtered targets totally 182 genes mRNAexpression profiles were analyzed in this studyTherewere 519interactions between target genes and organ locations Thenetworks of target genes tissuemRNAexpression profiles andcompounds of BSS are shown in Figure 5
10 Evidence-Based Complementary and Alternative Medicine
GO Analysis
688
79
89
899
88910111111
1012
915
1219
1820
2218
2820
34
61 48 49
59 46
56 48 60 55 56 70 70 62 54 47 50 51 63
62 110
62 101
50 64
62 58
117 53
136 233
0 10 20 30 40 50response to fatty acid
circadian rhythmresponse to nutrient
glucose transportcellular response to lipopolysaccharide
steroid hormone mediated signaling pathwayresponse to ethanol
response to hydrogen peroxidepositive regulation of protein kinase B signaling
positive regulation of MAPK cascaderesponse to nicotine
response to coldcholesterol metabolic process
platelet activationpositive regulation of ERK1 and ERK2 cascade
response to lipopolysaccharidelipid metabolic process
response to estradioltranscription initiation from RNA polymerase II promoter
cell-cell signalingresponse to glucocorticoid
positive regulation of transcription DNA-templatedcell proliferation
positive regulation of cell proliferationoxidation-reduction process
positive regulation of transcription from RNA polymerase II promoter
Figure 2GOanalysis 30 enriched biological process (BP) of gene ontology (GO) terms sorted by119875 valuelt 001 and gene counts are displayedThe 119910-axis represents enriched biological process (BP) terms for the target genes and the 119909-axis shows gene counts and minus log 10 (119875 value)
As a result 159 of 182 target genes displayed beyond aver-agemRNAexpression in relevant organ tissues such as uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smoothmuscle andwhole bloodThe rest of23 genes of 182 targets did not display above average mRNAexpression in above organ tissues for example gamma-aminobutyric acid receptor subunit alpha-6 (GABRA6) andcoagulation factor X (F10)
Nevertheless most genes of 159 demonstrated highexpression patterns in several organs of parturition relatedtissues at the same time In detail 60 genes showed mostsignificantmRNA expression in the uterus andor uterus cor-pus group 130 for placenta andor fetus 86 for hypothalamusandor pituitary 82 for smoothmuscle 80 for pituitary and 81for whole blood Besides 30 of 159 genes showed expression
in all of 6 groups For instance muscarinic acetylcholinereceptor M2 (CHRM2) neuronal acetylcholine receptorsubunit 120572-2 (CHRNA2) gamma-aminobutyric acid receptorsubunit alpha-3 (GABRA3) NO synthase inducible (NOS2)cGMP-inhibited 3101584051015840-cyclic phosphodiesterase A (PDE3A)and sodium-dependent dopamine transporter (SLC6A3)recorded beyond average mRNA expression in all six groupsFurthermore 79 of targets were expressed in two or moreorgan tissues which suggests that those organs and targetgenes of BSS are closely correlated
4 Discussion
In this study network pharmacology method with DL OBCaco-2 and LR evaluation multiple drug-target prediction
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
(C52 DL = 027 OB = 6595 Caco-2 = 088) was confirmedto have a protective effect on injured human umbilical veinendothelial cells [36] and myricanone (C48 DL = 051 OB= 5761 Caco-2 = 067) was found to best inhibit mouse skintumor progression [37]
32 Target Fishing The 65 active compounds interact with185 target proteins as shown in Table 2 in other wordson average each compound on average interacts with 285target proteinsThis result confirms the polypharmacologicalcharacter of oriental medicine and demonstrates the syn-ergistic effects of multiple compounds on multiple targets[38] Different compounds in CG and DG can directly affectcommon targets for example the target protein ldquocalmodulin(CALM1)rdquo interacts with crysophanol fromCG and coniferylferulate from DG at the same time which implies thesynergetic or cumulative effects of herbal medicine
33 GO Analysis 397 biological process terms with 119875 valuesof lt001 were sorted using the functional annotation chart ofthe DAVID 68 Gene Functional Classification Tool basedon 185 filtered target genes and 119875 values were adjustedusing the Benjamini-Hochberg method 30 enriched GO BPterms extracted by 119875 value and gene counts are displayed inFigure 2 It is meaningful that most of the target genes aresignificantly related to the various BP involved in parturitionFor instance 30 extracted GO BP terms include ldquoMAPKsignaling pathwaysrdquo ldquosteroid hormone mediated signalingpathwayrdquo ldquoresponse to glucocorticoidrdquo ldquoresponse to estra-diolrdquo and ldquopositive regulation of ERK1 and ERK2 cascaderdquoldquoMAPK signaling pathwaysrdquo were reported to be activatedin human uterine cervical ripening during parturition [39]ldquoSteroid hormone mediated signaling pathwayrdquo is highly
related to parturition process as estrogen and progesteroneplay important roles in pregnancy and parturition andestrogen induceS the principal stimulatory myometrial con-tractility [40] Also estradiol takes key place in parturitionprocess [41] It was identified that increased ERK activationis observed at the onset of labor and it promotes myometrialcontractility and development of parturition [42 43] To sumup the target genes of BSS are highly associated with thebiological process (BP) of parturition
34 Network Construction and Analysis Network analysis isan efficient tool for visualizing and understanding multipletargeted drug actions and demonstrates drug actions withinthe context of the whole genome [44 45] For a better insightof therapeutic impacts H-C-T and T-P networks were con-structed and displayed in Figures 3 and 4 respectively In theH-C-T network nodes represent herb names compoundsand targets Also in the T-P network circular nodes representtargets and triangle nodes represent pathways Besides nodesize is relative to the degree and edges show interactionsbetween nodes
H-C-T network confirmed that there were 739 interac-tions between 185 targets and 65 active compounds of CGand DG oleic acid (C48 degree = 42) with the highestnumber of interactions with targets followed by succinicacid (C63 degree = 40) and stigmasterol (C62 degree =37) It shows that single molecules target multiple recep-tors [46] Also some compounds from CG and DG werefound to share common targets Likewise prostaglandinGH synthase 2 (PTGS2 degree = 56) displayed the mostaffinitive connections with compounds followed by gamma-aminobutyric acid receptor subunit alpha-1 (GABRA1 degree= 48) prostaglandin GH synthase 1 (PTGS1 degree = 37)
6 Evidence-Based Complementary and Alternative Medicine
Table 2 Related targets of potential compounds in BSS
Evidence-Based Complementary and Alternative Medicine 9
Table 2 Continued
UniProt ID Target name Gene NameP19793 Retinoic acid receptor RXR-alpha RXRAO00767 Acyl-CoA desaturase SCDQ14524 Sodium channel protein type 5 subunit alpha SCN5AP31040 Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial SDHAP16109 P-selectin SELPP05121 Plasminogen activator inhibitor 1 SERPINE1P14410 Sucrase-isomaltase intestinal SIO76082 Solute carrier family 22 member 5 SLC22A5Q9UBX3 Mitochondrial dicarboxylate carrier SLC25A10P11168 Solute carrier family 2 facilitated glucose transporter member 2 SLC2A2P23975 Sodium-dependent noradrenaline transporter SLC6A2Q01959 Sodium-dependent dopamine transporter SLC6A3P31645 Sodium-dependent serotonin transporter SLC6A4P35610 Sterol O-acyltransferase 1 SOAT1P00441 Superoxide dismutase [Cu-Zn] SOD1P08047 Transcription factor Sp1 SP1P12931 Proto-oncogene tyrosine-protein kinase SRC SRCP36956 Sterol regulatory element-binding protein 1 SREBF1Q12772 Sterol regulatory element-binding protein 2 SREBF2Q9P2R7 Succinyl-CoA ligase [ADP-forming] beta-chain mitochondrial SUCLA2Q99973 Telomerase protein component 1 TEP1P01375 Tumor necrosis factor TNFQ16881 Thioredoxin reductase cytoplasmic TXNRD1P55851 Mitochondrial uncoupling protein 2 UCP2P55916 Mitochondrial uncoupling protein 3 UCP3
and muscarinic acetylcholine receptor M1 (CHRM1 degree= 37) Except for C60 (PLA2G1B degree = 1) the rest ofthe 64 active compounds are connected with more thanone target likewise 73 (395) target genes out of 185interactedwithmore than one compoundThis result demon-strates the multicompounds and multitarget properties ofherbal compounds and there was a report that compoundswith multiple targets could have greater therapeutic efficacy[47]
In addition the top 40 pathways were extracted basedon gene counts and 119875 value (lt005) and 119875 value wasadjusted by Benjamini-Hochbergmethod T-P network usingrelevant targets of herbal compounds is demonstrated inFigure 4 There were 485 interactions between the top 40pathways and 135 of 185 target genes ldquoMetabolic pathwaysrdquo(degree = 49) and ldquoneuroactive ligand-receptor interactionpathwayrdquo (degree = 32) had the highest and the secondhighest numbers of connections with the targets followedby ldquocalcium signalingrdquo (degree = 21) ldquocAMP signaling path-wayrdquo (degree = 17) and ldquocGMP PKG signaling pathwayrdquo(degree = 15) These are compelling results that parturitionprocesses are the complex hormone interactions and it iswell known that calcium signals within the myometrium arepivotal for uterine contractions [48] In the same mannersome target genes demonstrated higher degree centralitywith top 40 pathways namely PI3-kinase subunit gamma(PIK3CG degree = 23) cAMP-dependent protein kinase
catalytic subunit alpha (PRKACA degree = 20) proteinkinase C beta type (PRKCB degree = 18) and calmodulin(CALM1 degree = 11) We can confirm the same result in theprevious researches For instance PI3-kinase subunit gammaplays the key role in regulating cAMP calcium cyclingand beta-adrenergic signaling [49] Moreover during thelabor calmodulin-calcium complex activates myosin light-chain kinase which causes the generation of ATPase activityeventually uterine contraction is promoted [50]
H-C-T network explains the multitarget multicom-pounds properties and accumulates effect of herbalmedicines and T-P network shows that target genes of BSSare highly related to the pathway associated with parturi-tion process
35 Target Organ Location Map It is important to confirmthe tissue mRNA expression profiles of the target genes atthe organ level to identify the effects of BSS on parturi-tion Since there was no mRNA expression information inBioGPS of muscarinic acetylcholine receptor M1 (CHRM1)putative beta-glucuronidase-like protein SMA3 (GUSBP1)and retinol-binding protein 2 (RBP2) excluding these 3targets from 185 filtered targets totally 182 genes mRNAexpression profiles were analyzed in this studyTherewere 519interactions between target genes and organ locations Thenetworks of target genes tissuemRNAexpression profiles andcompounds of BSS are shown in Figure 5
10 Evidence-Based Complementary and Alternative Medicine
GO Analysis
688
79
89
899
88910111111
1012
915
1219
1820
2218
2820
34
61 48 49
59 46
56 48 60 55 56 70 70 62 54 47 50 51 63
62 110
62 101
50 64
62 58
117 53
136 233
0 10 20 30 40 50response to fatty acid
circadian rhythmresponse to nutrient
glucose transportcellular response to lipopolysaccharide
steroid hormone mediated signaling pathwayresponse to ethanol
response to hydrogen peroxidepositive regulation of protein kinase B signaling
positive regulation of MAPK cascaderesponse to nicotine
response to coldcholesterol metabolic process
platelet activationpositive regulation of ERK1 and ERK2 cascade
response to lipopolysaccharidelipid metabolic process
response to estradioltranscription initiation from RNA polymerase II promoter
cell-cell signalingresponse to glucocorticoid
positive regulation of transcription DNA-templatedcell proliferation
positive regulation of cell proliferationoxidation-reduction process
positive regulation of transcription from RNA polymerase II promoter
Figure 2GOanalysis 30 enriched biological process (BP) of gene ontology (GO) terms sorted by119875 valuelt 001 and gene counts are displayedThe 119910-axis represents enriched biological process (BP) terms for the target genes and the 119909-axis shows gene counts and minus log 10 (119875 value)
As a result 159 of 182 target genes displayed beyond aver-agemRNAexpression in relevant organ tissues such as uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smoothmuscle andwhole bloodThe rest of23 genes of 182 targets did not display above average mRNAexpression in above organ tissues for example gamma-aminobutyric acid receptor subunit alpha-6 (GABRA6) andcoagulation factor X (F10)
Nevertheless most genes of 159 demonstrated highexpression patterns in several organs of parturition relatedtissues at the same time In detail 60 genes showed mostsignificantmRNA expression in the uterus andor uterus cor-pus group 130 for placenta andor fetus 86 for hypothalamusandor pituitary 82 for smoothmuscle 80 for pituitary and 81for whole blood Besides 30 of 159 genes showed expression
in all of 6 groups For instance muscarinic acetylcholinereceptor M2 (CHRM2) neuronal acetylcholine receptorsubunit 120572-2 (CHRNA2) gamma-aminobutyric acid receptorsubunit alpha-3 (GABRA3) NO synthase inducible (NOS2)cGMP-inhibited 3101584051015840-cyclic phosphodiesterase A (PDE3A)and sodium-dependent dopamine transporter (SLC6A3)recorded beyond average mRNA expression in all six groupsFurthermore 79 of targets were expressed in two or moreorgan tissues which suggests that those organs and targetgenes of BSS are closely correlated
4 Discussion
In this study network pharmacology method with DL OBCaco-2 and LR evaluation multiple drug-target prediction
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
Evidence-Based Complementary and Alternative Medicine 9
Table 2 Continued
UniProt ID Target name Gene NameP19793 Retinoic acid receptor RXR-alpha RXRAO00767 Acyl-CoA desaturase SCDQ14524 Sodium channel protein type 5 subunit alpha SCN5AP31040 Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial SDHAP16109 P-selectin SELPP05121 Plasminogen activator inhibitor 1 SERPINE1P14410 Sucrase-isomaltase intestinal SIO76082 Solute carrier family 22 member 5 SLC22A5Q9UBX3 Mitochondrial dicarboxylate carrier SLC25A10P11168 Solute carrier family 2 facilitated glucose transporter member 2 SLC2A2P23975 Sodium-dependent noradrenaline transporter SLC6A2Q01959 Sodium-dependent dopamine transporter SLC6A3P31645 Sodium-dependent serotonin transporter SLC6A4P35610 Sterol O-acyltransferase 1 SOAT1P00441 Superoxide dismutase [Cu-Zn] SOD1P08047 Transcription factor Sp1 SP1P12931 Proto-oncogene tyrosine-protein kinase SRC SRCP36956 Sterol regulatory element-binding protein 1 SREBF1Q12772 Sterol regulatory element-binding protein 2 SREBF2Q9P2R7 Succinyl-CoA ligase [ADP-forming] beta-chain mitochondrial SUCLA2Q99973 Telomerase protein component 1 TEP1P01375 Tumor necrosis factor TNFQ16881 Thioredoxin reductase cytoplasmic TXNRD1P55851 Mitochondrial uncoupling protein 2 UCP2P55916 Mitochondrial uncoupling protein 3 UCP3
and muscarinic acetylcholine receptor M1 (CHRM1 degree= 37) Except for C60 (PLA2G1B degree = 1) the rest ofthe 64 active compounds are connected with more thanone target likewise 73 (395) target genes out of 185interactedwithmore than one compoundThis result demon-strates the multicompounds and multitarget properties ofherbal compounds and there was a report that compoundswith multiple targets could have greater therapeutic efficacy[47]
In addition the top 40 pathways were extracted basedon gene counts and 119875 value (lt005) and 119875 value wasadjusted by Benjamini-Hochbergmethod T-P network usingrelevant targets of herbal compounds is demonstrated inFigure 4 There were 485 interactions between the top 40pathways and 135 of 185 target genes ldquoMetabolic pathwaysrdquo(degree = 49) and ldquoneuroactive ligand-receptor interactionpathwayrdquo (degree = 32) had the highest and the secondhighest numbers of connections with the targets followedby ldquocalcium signalingrdquo (degree = 21) ldquocAMP signaling path-wayrdquo (degree = 17) and ldquocGMP PKG signaling pathwayrdquo(degree = 15) These are compelling results that parturitionprocesses are the complex hormone interactions and it iswell known that calcium signals within the myometrium arepivotal for uterine contractions [48] In the same mannersome target genes demonstrated higher degree centralitywith top 40 pathways namely PI3-kinase subunit gamma(PIK3CG degree = 23) cAMP-dependent protein kinase
catalytic subunit alpha (PRKACA degree = 20) proteinkinase C beta type (PRKCB degree = 18) and calmodulin(CALM1 degree = 11) We can confirm the same result in theprevious researches For instance PI3-kinase subunit gammaplays the key role in regulating cAMP calcium cyclingand beta-adrenergic signaling [49] Moreover during thelabor calmodulin-calcium complex activates myosin light-chain kinase which causes the generation of ATPase activityeventually uterine contraction is promoted [50]
H-C-T network explains the multitarget multicom-pounds properties and accumulates effect of herbalmedicines and T-P network shows that target genes of BSSare highly related to the pathway associated with parturi-tion process
35 Target Organ Location Map It is important to confirmthe tissue mRNA expression profiles of the target genes atthe organ level to identify the effects of BSS on parturi-tion Since there was no mRNA expression information inBioGPS of muscarinic acetylcholine receptor M1 (CHRM1)putative beta-glucuronidase-like protein SMA3 (GUSBP1)and retinol-binding protein 2 (RBP2) excluding these 3targets from 185 filtered targets totally 182 genes mRNAexpression profiles were analyzed in this studyTherewere 519interactions between target genes and organ locations Thenetworks of target genes tissuemRNAexpression profiles andcompounds of BSS are shown in Figure 5
10 Evidence-Based Complementary and Alternative Medicine
GO Analysis
688
79
89
899
88910111111
1012
915
1219
1820
2218
2820
34
61 48 49
59 46
56 48 60 55 56 70 70 62 54 47 50 51 63
62 110
62 101
50 64
62 58
117 53
136 233
0 10 20 30 40 50response to fatty acid
circadian rhythmresponse to nutrient
glucose transportcellular response to lipopolysaccharide
steroid hormone mediated signaling pathwayresponse to ethanol
response to hydrogen peroxidepositive regulation of protein kinase B signaling
positive regulation of MAPK cascaderesponse to nicotine
response to coldcholesterol metabolic process
platelet activationpositive regulation of ERK1 and ERK2 cascade
response to lipopolysaccharidelipid metabolic process
response to estradioltranscription initiation from RNA polymerase II promoter
cell-cell signalingresponse to glucocorticoid
positive regulation of transcription DNA-templatedcell proliferation
positive regulation of cell proliferationoxidation-reduction process
positive regulation of transcription from RNA polymerase II promoter
Figure 2GOanalysis 30 enriched biological process (BP) of gene ontology (GO) terms sorted by119875 valuelt 001 and gene counts are displayedThe 119910-axis represents enriched biological process (BP) terms for the target genes and the 119909-axis shows gene counts and minus log 10 (119875 value)
As a result 159 of 182 target genes displayed beyond aver-agemRNAexpression in relevant organ tissues such as uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smoothmuscle andwhole bloodThe rest of23 genes of 182 targets did not display above average mRNAexpression in above organ tissues for example gamma-aminobutyric acid receptor subunit alpha-6 (GABRA6) andcoagulation factor X (F10)
Nevertheless most genes of 159 demonstrated highexpression patterns in several organs of parturition relatedtissues at the same time In detail 60 genes showed mostsignificantmRNA expression in the uterus andor uterus cor-pus group 130 for placenta andor fetus 86 for hypothalamusandor pituitary 82 for smoothmuscle 80 for pituitary and 81for whole blood Besides 30 of 159 genes showed expression
in all of 6 groups For instance muscarinic acetylcholinereceptor M2 (CHRM2) neuronal acetylcholine receptorsubunit 120572-2 (CHRNA2) gamma-aminobutyric acid receptorsubunit alpha-3 (GABRA3) NO synthase inducible (NOS2)cGMP-inhibited 3101584051015840-cyclic phosphodiesterase A (PDE3A)and sodium-dependent dopamine transporter (SLC6A3)recorded beyond average mRNA expression in all six groupsFurthermore 79 of targets were expressed in two or moreorgan tissues which suggests that those organs and targetgenes of BSS are closely correlated
4 Discussion
In this study network pharmacology method with DL OBCaco-2 and LR evaluation multiple drug-target prediction
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
Evidence-Based Complementary and Alternative Medicine 9
Table 2 Continued
UniProt ID Target name Gene NameP19793 Retinoic acid receptor RXR-alpha RXRAO00767 Acyl-CoA desaturase SCDQ14524 Sodium channel protein type 5 subunit alpha SCN5AP31040 Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial SDHAP16109 P-selectin SELPP05121 Plasminogen activator inhibitor 1 SERPINE1P14410 Sucrase-isomaltase intestinal SIO76082 Solute carrier family 22 member 5 SLC22A5Q9UBX3 Mitochondrial dicarboxylate carrier SLC25A10P11168 Solute carrier family 2 facilitated glucose transporter member 2 SLC2A2P23975 Sodium-dependent noradrenaline transporter SLC6A2Q01959 Sodium-dependent dopamine transporter SLC6A3P31645 Sodium-dependent serotonin transporter SLC6A4P35610 Sterol O-acyltransferase 1 SOAT1P00441 Superoxide dismutase [Cu-Zn] SOD1P08047 Transcription factor Sp1 SP1P12931 Proto-oncogene tyrosine-protein kinase SRC SRCP36956 Sterol regulatory element-binding protein 1 SREBF1Q12772 Sterol regulatory element-binding protein 2 SREBF2Q9P2R7 Succinyl-CoA ligase [ADP-forming] beta-chain mitochondrial SUCLA2Q99973 Telomerase protein component 1 TEP1P01375 Tumor necrosis factor TNFQ16881 Thioredoxin reductase cytoplasmic TXNRD1P55851 Mitochondrial uncoupling protein 2 UCP2P55916 Mitochondrial uncoupling protein 3 UCP3
and muscarinic acetylcholine receptor M1 (CHRM1 degree= 37) Except for C60 (PLA2G1B degree = 1) the rest ofthe 64 active compounds are connected with more thanone target likewise 73 (395) target genes out of 185interactedwithmore than one compoundThis result demon-strates the multicompounds and multitarget properties ofherbal compounds and there was a report that compoundswith multiple targets could have greater therapeutic efficacy[47]
In addition the top 40 pathways were extracted basedon gene counts and 119875 value (lt005) and 119875 value wasadjusted by Benjamini-Hochbergmethod T-P network usingrelevant targets of herbal compounds is demonstrated inFigure 4 There were 485 interactions between the top 40pathways and 135 of 185 target genes ldquoMetabolic pathwaysrdquo(degree = 49) and ldquoneuroactive ligand-receptor interactionpathwayrdquo (degree = 32) had the highest and the secondhighest numbers of connections with the targets followedby ldquocalcium signalingrdquo (degree = 21) ldquocAMP signaling path-wayrdquo (degree = 17) and ldquocGMP PKG signaling pathwayrdquo(degree = 15) These are compelling results that parturitionprocesses are the complex hormone interactions and it iswell known that calcium signals within the myometrium arepivotal for uterine contractions [48] In the same mannersome target genes demonstrated higher degree centralitywith top 40 pathways namely PI3-kinase subunit gamma(PIK3CG degree = 23) cAMP-dependent protein kinase
catalytic subunit alpha (PRKACA degree = 20) proteinkinase C beta type (PRKCB degree = 18) and calmodulin(CALM1 degree = 11) We can confirm the same result in theprevious researches For instance PI3-kinase subunit gammaplays the key role in regulating cAMP calcium cyclingand beta-adrenergic signaling [49] Moreover during thelabor calmodulin-calcium complex activates myosin light-chain kinase which causes the generation of ATPase activityeventually uterine contraction is promoted [50]
H-C-T network explains the multitarget multicom-pounds properties and accumulates effect of herbalmedicines and T-P network shows that target genes of BSSare highly related to the pathway associated with parturi-tion process
35 Target Organ Location Map It is important to confirmthe tissue mRNA expression profiles of the target genes atthe organ level to identify the effects of BSS on parturi-tion Since there was no mRNA expression information inBioGPS of muscarinic acetylcholine receptor M1 (CHRM1)putative beta-glucuronidase-like protein SMA3 (GUSBP1)and retinol-binding protein 2 (RBP2) excluding these 3targets from 185 filtered targets totally 182 genes mRNAexpression profiles were analyzed in this studyTherewere 519interactions between target genes and organ locations Thenetworks of target genes tissuemRNAexpression profiles andcompounds of BSS are shown in Figure 5
10 Evidence-Based Complementary and Alternative Medicine
GO Analysis
688
79
89
899
88910111111
1012
915
1219
1820
2218
2820
34
61 48 49
59 46
56 48 60 55 56 70 70 62 54 47 50 51 63
62 110
62 101
50 64
62 58
117 53
136 233
0 10 20 30 40 50response to fatty acid
circadian rhythmresponse to nutrient
glucose transportcellular response to lipopolysaccharide
steroid hormone mediated signaling pathwayresponse to ethanol
response to hydrogen peroxidepositive regulation of protein kinase B signaling
positive regulation of MAPK cascaderesponse to nicotine
response to coldcholesterol metabolic process
platelet activationpositive regulation of ERK1 and ERK2 cascade
response to lipopolysaccharidelipid metabolic process
response to estradioltranscription initiation from RNA polymerase II promoter
cell-cell signalingresponse to glucocorticoid
positive regulation of transcription DNA-templatedcell proliferation
positive regulation of cell proliferationoxidation-reduction process
positive regulation of transcription from RNA polymerase II promoter
Figure 2GOanalysis 30 enriched biological process (BP) of gene ontology (GO) terms sorted by119875 valuelt 001 and gene counts are displayedThe 119910-axis represents enriched biological process (BP) terms for the target genes and the 119909-axis shows gene counts and minus log 10 (119875 value)
As a result 159 of 182 target genes displayed beyond aver-agemRNAexpression in relevant organ tissues such as uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smoothmuscle andwhole bloodThe rest of23 genes of 182 targets did not display above average mRNAexpression in above organ tissues for example gamma-aminobutyric acid receptor subunit alpha-6 (GABRA6) andcoagulation factor X (F10)
Nevertheless most genes of 159 demonstrated highexpression patterns in several organs of parturition relatedtissues at the same time In detail 60 genes showed mostsignificantmRNA expression in the uterus andor uterus cor-pus group 130 for placenta andor fetus 86 for hypothalamusandor pituitary 82 for smoothmuscle 80 for pituitary and 81for whole blood Besides 30 of 159 genes showed expression
in all of 6 groups For instance muscarinic acetylcholinereceptor M2 (CHRM2) neuronal acetylcholine receptorsubunit 120572-2 (CHRNA2) gamma-aminobutyric acid receptorsubunit alpha-3 (GABRA3) NO synthase inducible (NOS2)cGMP-inhibited 3101584051015840-cyclic phosphodiesterase A (PDE3A)and sodium-dependent dopamine transporter (SLC6A3)recorded beyond average mRNA expression in all six groupsFurthermore 79 of targets were expressed in two or moreorgan tissues which suggests that those organs and targetgenes of BSS are closely correlated
4 Discussion
In this study network pharmacology method with DL OBCaco-2 and LR evaluation multiple drug-target prediction
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
Evidence-Based Complementary and Alternative Medicine 9
Table 2 Continued
UniProt ID Target name Gene NameP19793 Retinoic acid receptor RXR-alpha RXRAO00767 Acyl-CoA desaturase SCDQ14524 Sodium channel protein type 5 subunit alpha SCN5AP31040 Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial SDHAP16109 P-selectin SELPP05121 Plasminogen activator inhibitor 1 SERPINE1P14410 Sucrase-isomaltase intestinal SIO76082 Solute carrier family 22 member 5 SLC22A5Q9UBX3 Mitochondrial dicarboxylate carrier SLC25A10P11168 Solute carrier family 2 facilitated glucose transporter member 2 SLC2A2P23975 Sodium-dependent noradrenaline transporter SLC6A2Q01959 Sodium-dependent dopamine transporter SLC6A3P31645 Sodium-dependent serotonin transporter SLC6A4P35610 Sterol O-acyltransferase 1 SOAT1P00441 Superoxide dismutase [Cu-Zn] SOD1P08047 Transcription factor Sp1 SP1P12931 Proto-oncogene tyrosine-protein kinase SRC SRCP36956 Sterol regulatory element-binding protein 1 SREBF1Q12772 Sterol regulatory element-binding protein 2 SREBF2Q9P2R7 Succinyl-CoA ligase [ADP-forming] beta-chain mitochondrial SUCLA2Q99973 Telomerase protein component 1 TEP1P01375 Tumor necrosis factor TNFQ16881 Thioredoxin reductase cytoplasmic TXNRD1P55851 Mitochondrial uncoupling protein 2 UCP2P55916 Mitochondrial uncoupling protein 3 UCP3
and muscarinic acetylcholine receptor M1 (CHRM1 degree= 37) Except for C60 (PLA2G1B degree = 1) the rest ofthe 64 active compounds are connected with more thanone target likewise 73 (395) target genes out of 185interactedwithmore than one compoundThis result demon-strates the multicompounds and multitarget properties ofherbal compounds and there was a report that compoundswith multiple targets could have greater therapeutic efficacy[47]
In addition the top 40 pathways were extracted basedon gene counts and 119875 value (lt005) and 119875 value wasadjusted by Benjamini-Hochbergmethod T-P network usingrelevant targets of herbal compounds is demonstrated inFigure 4 There were 485 interactions between the top 40pathways and 135 of 185 target genes ldquoMetabolic pathwaysrdquo(degree = 49) and ldquoneuroactive ligand-receptor interactionpathwayrdquo (degree = 32) had the highest and the secondhighest numbers of connections with the targets followedby ldquocalcium signalingrdquo (degree = 21) ldquocAMP signaling path-wayrdquo (degree = 17) and ldquocGMP PKG signaling pathwayrdquo(degree = 15) These are compelling results that parturitionprocesses are the complex hormone interactions and it iswell known that calcium signals within the myometrium arepivotal for uterine contractions [48] In the same mannersome target genes demonstrated higher degree centralitywith top 40 pathways namely PI3-kinase subunit gamma(PIK3CG degree = 23) cAMP-dependent protein kinase
catalytic subunit alpha (PRKACA degree = 20) proteinkinase C beta type (PRKCB degree = 18) and calmodulin(CALM1 degree = 11) We can confirm the same result in theprevious researches For instance PI3-kinase subunit gammaplays the key role in regulating cAMP calcium cyclingand beta-adrenergic signaling [49] Moreover during thelabor calmodulin-calcium complex activates myosin light-chain kinase which causes the generation of ATPase activityeventually uterine contraction is promoted [50]
H-C-T network explains the multitarget multicom-pounds properties and accumulates effect of herbalmedicines and T-P network shows that target genes of BSSare highly related to the pathway associated with parturi-tion process
35 Target Organ Location Map It is important to confirmthe tissue mRNA expression profiles of the target genes atthe organ level to identify the effects of BSS on parturi-tion Since there was no mRNA expression information inBioGPS of muscarinic acetylcholine receptor M1 (CHRM1)putative beta-glucuronidase-like protein SMA3 (GUSBP1)and retinol-binding protein 2 (RBP2) excluding these 3targets from 185 filtered targets totally 182 genes mRNAexpression profiles were analyzed in this studyTherewere 519interactions between target genes and organ locations Thenetworks of target genes tissuemRNAexpression profiles andcompounds of BSS are shown in Figure 5
10 Evidence-Based Complementary and Alternative Medicine
GO Analysis
688
79
89
899
88910111111
1012
915
1219
1820
2218
2820
34
61 48 49
59 46
56 48 60 55 56 70 70 62 54 47 50 51 63
62 110
62 101
50 64
62 58
117 53
136 233
0 10 20 30 40 50response to fatty acid
circadian rhythmresponse to nutrient
glucose transportcellular response to lipopolysaccharide
steroid hormone mediated signaling pathwayresponse to ethanol
response to hydrogen peroxidepositive regulation of protein kinase B signaling
positive regulation of MAPK cascaderesponse to nicotine
response to coldcholesterol metabolic process
platelet activationpositive regulation of ERK1 and ERK2 cascade
response to lipopolysaccharidelipid metabolic process
response to estradioltranscription initiation from RNA polymerase II promoter
cell-cell signalingresponse to glucocorticoid
positive regulation of transcription DNA-templatedcell proliferation
positive regulation of cell proliferationoxidation-reduction process
positive regulation of transcription from RNA polymerase II promoter
Figure 2GOanalysis 30 enriched biological process (BP) of gene ontology (GO) terms sorted by119875 valuelt 001 and gene counts are displayedThe 119910-axis represents enriched biological process (BP) terms for the target genes and the 119909-axis shows gene counts and minus log 10 (119875 value)
As a result 159 of 182 target genes displayed beyond aver-agemRNAexpression in relevant organ tissues such as uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smoothmuscle andwhole bloodThe rest of23 genes of 182 targets did not display above average mRNAexpression in above organ tissues for example gamma-aminobutyric acid receptor subunit alpha-6 (GABRA6) andcoagulation factor X (F10)
Nevertheless most genes of 159 demonstrated highexpression patterns in several organs of parturition relatedtissues at the same time In detail 60 genes showed mostsignificantmRNA expression in the uterus andor uterus cor-pus group 130 for placenta andor fetus 86 for hypothalamusandor pituitary 82 for smoothmuscle 80 for pituitary and 81for whole blood Besides 30 of 159 genes showed expression
in all of 6 groups For instance muscarinic acetylcholinereceptor M2 (CHRM2) neuronal acetylcholine receptorsubunit 120572-2 (CHRNA2) gamma-aminobutyric acid receptorsubunit alpha-3 (GABRA3) NO synthase inducible (NOS2)cGMP-inhibited 3101584051015840-cyclic phosphodiesterase A (PDE3A)and sodium-dependent dopamine transporter (SLC6A3)recorded beyond average mRNA expression in all six groupsFurthermore 79 of targets were expressed in two or moreorgan tissues which suggests that those organs and targetgenes of BSS are closely correlated
4 Discussion
In this study network pharmacology method with DL OBCaco-2 and LR evaluation multiple drug-target prediction
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
Evidence-Based Complementary and Alternative Medicine 9
Table 2 Continued
UniProt ID Target name Gene NameP19793 Retinoic acid receptor RXR-alpha RXRAO00767 Acyl-CoA desaturase SCDQ14524 Sodium channel protein type 5 subunit alpha SCN5AP31040 Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial SDHAP16109 P-selectin SELPP05121 Plasminogen activator inhibitor 1 SERPINE1P14410 Sucrase-isomaltase intestinal SIO76082 Solute carrier family 22 member 5 SLC22A5Q9UBX3 Mitochondrial dicarboxylate carrier SLC25A10P11168 Solute carrier family 2 facilitated glucose transporter member 2 SLC2A2P23975 Sodium-dependent noradrenaline transporter SLC6A2Q01959 Sodium-dependent dopamine transporter SLC6A3P31645 Sodium-dependent serotonin transporter SLC6A4P35610 Sterol O-acyltransferase 1 SOAT1P00441 Superoxide dismutase [Cu-Zn] SOD1P08047 Transcription factor Sp1 SP1P12931 Proto-oncogene tyrosine-protein kinase SRC SRCP36956 Sterol regulatory element-binding protein 1 SREBF1Q12772 Sterol regulatory element-binding protein 2 SREBF2Q9P2R7 Succinyl-CoA ligase [ADP-forming] beta-chain mitochondrial SUCLA2Q99973 Telomerase protein component 1 TEP1P01375 Tumor necrosis factor TNFQ16881 Thioredoxin reductase cytoplasmic TXNRD1P55851 Mitochondrial uncoupling protein 2 UCP2P55916 Mitochondrial uncoupling protein 3 UCP3
and muscarinic acetylcholine receptor M1 (CHRM1 degree= 37) Except for C60 (PLA2G1B degree = 1) the rest ofthe 64 active compounds are connected with more thanone target likewise 73 (395) target genes out of 185interactedwithmore than one compoundThis result demon-strates the multicompounds and multitarget properties ofherbal compounds and there was a report that compoundswith multiple targets could have greater therapeutic efficacy[47]
In addition the top 40 pathways were extracted basedon gene counts and 119875 value (lt005) and 119875 value wasadjusted by Benjamini-Hochbergmethod T-P network usingrelevant targets of herbal compounds is demonstrated inFigure 4 There were 485 interactions between the top 40pathways and 135 of 185 target genes ldquoMetabolic pathwaysrdquo(degree = 49) and ldquoneuroactive ligand-receptor interactionpathwayrdquo (degree = 32) had the highest and the secondhighest numbers of connections with the targets followedby ldquocalcium signalingrdquo (degree = 21) ldquocAMP signaling path-wayrdquo (degree = 17) and ldquocGMP PKG signaling pathwayrdquo(degree = 15) These are compelling results that parturitionprocesses are the complex hormone interactions and it iswell known that calcium signals within the myometrium arepivotal for uterine contractions [48] In the same mannersome target genes demonstrated higher degree centralitywith top 40 pathways namely PI3-kinase subunit gamma(PIK3CG degree = 23) cAMP-dependent protein kinase
catalytic subunit alpha (PRKACA degree = 20) proteinkinase C beta type (PRKCB degree = 18) and calmodulin(CALM1 degree = 11) We can confirm the same result in theprevious researches For instance PI3-kinase subunit gammaplays the key role in regulating cAMP calcium cyclingand beta-adrenergic signaling [49] Moreover during thelabor calmodulin-calcium complex activates myosin light-chain kinase which causes the generation of ATPase activityeventually uterine contraction is promoted [50]
H-C-T network explains the multitarget multicom-pounds properties and accumulates effect of herbalmedicines and T-P network shows that target genes of BSSare highly related to the pathway associated with parturi-tion process
35 Target Organ Location Map It is important to confirmthe tissue mRNA expression profiles of the target genes atthe organ level to identify the effects of BSS on parturi-tion Since there was no mRNA expression information inBioGPS of muscarinic acetylcholine receptor M1 (CHRM1)putative beta-glucuronidase-like protein SMA3 (GUSBP1)and retinol-binding protein 2 (RBP2) excluding these 3targets from 185 filtered targets totally 182 genes mRNAexpression profiles were analyzed in this studyTherewere 519interactions between target genes and organ locations Thenetworks of target genes tissuemRNAexpression profiles andcompounds of BSS are shown in Figure 5
10 Evidence-Based Complementary and Alternative Medicine
GO Analysis
688
79
89
899
88910111111
1012
915
1219
1820
2218
2820
34
61 48 49
59 46
56 48 60 55 56 70 70 62 54 47 50 51 63
62 110
62 101
50 64
62 58
117 53
136 233
0 10 20 30 40 50response to fatty acid
circadian rhythmresponse to nutrient
glucose transportcellular response to lipopolysaccharide
steroid hormone mediated signaling pathwayresponse to ethanol
response to hydrogen peroxidepositive regulation of protein kinase B signaling
positive regulation of MAPK cascaderesponse to nicotine
response to coldcholesterol metabolic process
platelet activationpositive regulation of ERK1 and ERK2 cascade
response to lipopolysaccharidelipid metabolic process
response to estradioltranscription initiation from RNA polymerase II promoter
cell-cell signalingresponse to glucocorticoid
positive regulation of transcription DNA-templatedcell proliferation
positive regulation of cell proliferationoxidation-reduction process
positive regulation of transcription from RNA polymerase II promoter
Figure 2GOanalysis 30 enriched biological process (BP) of gene ontology (GO) terms sorted by119875 valuelt 001 and gene counts are displayedThe 119910-axis represents enriched biological process (BP) terms for the target genes and the 119909-axis shows gene counts and minus log 10 (119875 value)
As a result 159 of 182 target genes displayed beyond aver-agemRNAexpression in relevant organ tissues such as uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smoothmuscle andwhole bloodThe rest of23 genes of 182 targets did not display above average mRNAexpression in above organ tissues for example gamma-aminobutyric acid receptor subunit alpha-6 (GABRA6) andcoagulation factor X (F10)
Nevertheless most genes of 159 demonstrated highexpression patterns in several organs of parturition relatedtissues at the same time In detail 60 genes showed mostsignificantmRNA expression in the uterus andor uterus cor-pus group 130 for placenta andor fetus 86 for hypothalamusandor pituitary 82 for smoothmuscle 80 for pituitary and 81for whole blood Besides 30 of 159 genes showed expression
in all of 6 groups For instance muscarinic acetylcholinereceptor M2 (CHRM2) neuronal acetylcholine receptorsubunit 120572-2 (CHRNA2) gamma-aminobutyric acid receptorsubunit alpha-3 (GABRA3) NO synthase inducible (NOS2)cGMP-inhibited 3101584051015840-cyclic phosphodiesterase A (PDE3A)and sodium-dependent dopamine transporter (SLC6A3)recorded beyond average mRNA expression in all six groupsFurthermore 79 of targets were expressed in two or moreorgan tissues which suggests that those organs and targetgenes of BSS are closely correlated
4 Discussion
In this study network pharmacology method with DL OBCaco-2 and LR evaluation multiple drug-target prediction
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
positive regulation of transcription from RNA polymerase II promoter
Figure 2GOanalysis 30 enriched biological process (BP) of gene ontology (GO) terms sorted by119875 valuelt 001 and gene counts are displayedThe 119910-axis represents enriched biological process (BP) terms for the target genes and the 119909-axis shows gene counts and minus log 10 (119875 value)
As a result 159 of 182 target genes displayed beyond aver-agemRNAexpression in relevant organ tissues such as uterusandor uterus corpus fetus andor placenta hypothalamusandor pituitary smoothmuscle andwhole bloodThe rest of23 genes of 182 targets did not display above average mRNAexpression in above organ tissues for example gamma-aminobutyric acid receptor subunit alpha-6 (GABRA6) andcoagulation factor X (F10)
Nevertheless most genes of 159 demonstrated highexpression patterns in several organs of parturition relatedtissues at the same time In detail 60 genes showed mostsignificantmRNA expression in the uterus andor uterus cor-pus group 130 for placenta andor fetus 86 for hypothalamusandor pituitary 82 for smoothmuscle 80 for pituitary and 81for whole blood Besides 30 of 159 genes showed expression
in all of 6 groups For instance muscarinic acetylcholinereceptor M2 (CHRM2) neuronal acetylcholine receptorsubunit 120572-2 (CHRNA2) gamma-aminobutyric acid receptorsubunit alpha-3 (GABRA3) NO synthase inducible (NOS2)cGMP-inhibited 3101584051015840-cyclic phosphodiesterase A (PDE3A)and sodium-dependent dopamine transporter (SLC6A3)recorded beyond average mRNA expression in all six groupsFurthermore 79 of targets were expressed in two or moreorgan tissues which suggests that those organs and targetgenes of BSS are closely correlated
4 Discussion
In this study network pharmacology method with DL OBCaco-2 and LR evaluation multiple drug-target prediction
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
Evidence-Based Complementary and Alternative Medicine 11
DPP4
CDK2
CHEK1
MAPK14
PIM1
CCNA2F7
PLG
ERBB2C41NTRK2
ESR2
CITED1
PON1
PLAU
SERPINE1RBP2
GSK3B
INS CETP
HP
C47
MOGAT2PPARA
APOB
PPARGC1A
ADH1C
EDNRA
HMGCR
ENPEP
PYY
GCGR
PDX1
PPARD
FABP1
SOAT1
TEP1
SOD1GAP43
MPO
UCP2
SLC2A2
SCDCAT
UCP3
C48
DNPEP
CCK
LPL
PDHX
NR3C1
C25
FOSL2
AMY2A
ACSL1
NR1I3
ADORA2A
ALAD
CYP1A2
PTEN
SREBF1FOS
AMY1A
PCYT1A
SLC22A5
PTPN1
G6PCACSL4
HK1
SREBF2 LCT
SI
COL7A1CHI3L1
C45
COL1A1
AKR1B1
GUSBP1
IL10
IL6PTGER3
C49C65
C64
C9C40
C20
C52
C59KDR
C22KCNH2
CYP1A1 NQO2
RAC1 CHATSELP
CA1
IGF2 GFAP
PKIAPDE3A
LTA4H
SCN5AHSP90AB1
C34
BTK
C13C17
ESR1
NCOA1
C46
F2
NOS2
C39
PRKACAPTGS2
PTGS1CG
C57
C62C14
RXRA
C50
C51
DG
NR3C2
TNF
GABRA1
IGHG1NOS3
NCOA2PGR
C61
SLC6A4
ADRA2B
ADRA2A
DRD2
HTR2A
ADRA1A
MAOA ADRB1
SLC6A3
CTRB1
MAOB
F10
ADRA2C
ADRB2
PIK3CG
CALM1
AR
PRKCB
ADRA1D
C4
C8
C27
C21
C2
C43
C56
C32
C58 C18C36
C15
C35
C28
C26
C23
C5
C38
C54
CHRM2
CHRM1
C3
C16 C44
CHRM3
C55
C11
C19C10
GRIA2 ADRA1B
C31
C7
C30
C33
C6
C53
PLA2G1B
ADA
PNP
SP1
AHCYL1
C60
C24
ENPP7
GABRA4DRD1
GABRA2
GABRA3
GABRA5
GABRA6
CHRNA2
SLC6A2
C1
CHRNA7
C12C29
C42
C37
KCNMA1
LEPREL2
P4HA2MAT1A
OATABAT
GIG18
ME3
PRSS3
GLRA1
HSD17B6
ALDH5A1
OXCT2
OTC
ADH1A
CASP3
PKM2
SDHA
GOT2
SLC25A10
BCHEME2
AGXT
SUCLA2 GOT1MAT2A
LEPRE1 BCL2
RHO
C63
ATP5B
ADH1B
CTSD
SRC
PPARG
ADSS
CDC25B GLUD1
NOS1
TXNRD1
Figure 3 H-C-T network herb-compound-target (H-C-T) network demonstrated multicompound multitarget property of BSS In thisnetwork red and blue nodes represent herbs green nodes show compounds and pink nodes indicate targets and node size is relative to thedegree and edges demonstrate interactions between nodes
network analysis and relevant organ location mapping wasused to explain the targets of BSS in relation to the parturitionprocess There is no denying that network based analysis ispowerful approach for identifying the actions of multitarget-ing herbal medicines at the systems level and our study showstarget genes of BSS are strongly connected to parturitionrelated pathways biological processes and organs It wasconfirmed that 98 of the active compounds of BSS wereinteracted with more than two targets and 395 of thetargets related to more than one compound The synergeticmultitarget properties of BSS were visualized but furtherdiscussion about differentiated drug action based on degreecentrality and simultaneous targeting effect of more than onecompound is required [51] Also detailed potential pathwaysof BSS should be explored deeply in the future
Similar findings were identified in a few RCT researchesin China that using BSS in induction of labor can reduce
the delivery time the amount of bleeding and the residualrate of placenta [52 53] In addition BSS targets six genes ofGABA receptor and NOS which was reported to be relatedoxytocin neurons at the time of parturition in rats [54] AlsoBSS targets NOS and NO (nitric oxide) which are involved inthe regulation of uterine contractility during pregnancy andis a key factor for the onset of labor [55] and iNOS (induciblenitric oxide synthase) can be upregulated accordantly bysimilar inflammatory mediators during ripening [11]
In fact rather than DG Angelicae Gigantis Radix (Dang-gwi AGR) grows naturally in Korea for that reason thecombination of AGR and CG is commonly used as BSSin Korea Instead DG is named as Chinese Danggwi foraccurate classification in Korea Several studies have shownAGR is differs from DG in terms of its main active con-stituents and genetic form AGR is mainly composed of
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
12 Evidence-Based Complementary and Alternative Medicine
SDHA
KDR
ADSSALAD
ADH1A
HK1
ENPP7ADH1C
Carbohydratedigestionandabsorption
NonalcoholicfattyliverdiseaseNAFLD
Insulinsignalingpathway
PRKACA
CHAT
cAMPsignalingpathway
CALM1
PRKCB
SLC6A3PLA2G1B
Cholinergicsynapse
Regulationoflipolysisinadipocytes
NOS2
PIK3CG
NOS3
RAC1 SRC
CASP3
GSK3B
VEGFsignalingpathway
HIF1signalingpathway
PPARA
NR3C1
Calciumsignalingpathway
Salivarysecretion
ADRB2
ADORA2ACHRM2
ADRB1
Vascularsmoothmusclecontraction
SOD1 Dopaminergicsynapse
PTGS2
ATP5BPDHX
LCT
HSD17B6
AMY2A
SI
ACSL4
AMY1A
ACSL1
ADA
cGMPPKGsignalingpathwayPTGER3MAPK14
BCL2KCNMA1ADRA1A
FOS
EDNRA
ADRA1B
CHRM3
ADRA2C
ADRA1D
ADRA2B
ADRA2A
GABRA1
GABRA4
GABRA6 GABRA3
GABRA5
Nicotineaddiction
DRD1
Serotonergicsynapse
NOS1
PTGS1
Retrogradeendocannabinoidsignaling
CHRNA7Amphetamineaddiction
CHRNA2
PDE3A
GLRA1
PLG
CHRM1
F2 GCGR
PRSS3
Neuroactiveligandreceptorinteraction
PNP
PCYT1A
Metabolicpathways
ADH1B
CYP1A2
CAT
MAT1AALDH5A1
Arginineandprolinemetabolism
ABATOAT
GOT1GABRA2LTA4H
PON1
CYP1A1P4HA2GOT2
Morphineaddiction
DRD2HTR2A
MAOB
MAOA GRIA2
GABAergicsynapseSLC6A4
ME2
GLUD1
Carbonmetabolism AHCYL1
AKR1B1
OTC
MAT2A SUCLA2
AGXT
ME3
FABP1
SREBF1
PTPN1
G6PC
HMGCR
SLC2A2
PPARsignalingpathway
CDK2CCNA2
ESR2 AR
ESR1
HSP90AB1
PPARG
PPARD
RXRA
LPLPPARGC1A
SCD
SCN5A
Sphingolipidsignalingpathway
Pertussis
Adrenergicsignalingincardiomyocytes
CTSD
ERBB2
Amoebiasis
TNF
InsulinresistanceNCOA2
Proteoglycansincancer
IGF2
INS
NCOA1PLAU
yroidhormonesignalingpathway
PathwaysincancerProlactinsignalingpathway COL1A1
IL6
EstrogensignalingpathwaySERPINE1
PGRAMPKsignalingpathway
IL10Prostatecancer
SP1
PTEN
HepatitisB
CDC25B
ChagasdiseaseAmericantrypanosomiasis
Progesteronemediatedoocytematuration
AmyotrophiclateralsclerosisALS
Figure 4 T-P network in target-pathway (T-P) network circular nodes represent compounds and triangles indicate pathways Node size isrelative to the degree and edges demonstrate interactions between nodes
water soluble polysaccharide but coumarin which is liposol-uble including nodakenin (1) peucedanone (2) marmesin(3) decursinol (4) 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-enyl) coumarin (5) demethylsuberosin (6) decursin (7)decursinol angelate (8) and isoimperatorin (9) [56] Ofthese decursin and its isomer decursinol angelate have beenreported to be the active compounds in AGR [57] It wasidentified in the experimental studies that AGR and DGact via different mechanisms in the cardiovascular centralnervous system and anticancer activity but both have similarpharmacological effects [57] Since the compositions of DGand AGR differ further study on BSS with AGR is requiredCurrently BSS is commonly prescribed to treat cerebravascular and cardiovascular diseases in China [33] but inKorea BSS is widely applied in obstetrics
The similarity between cervical ripening during partu-rition and inflammatory reaction has been pointed out inearlier studies this has been attributed to the inductionof leukocyte migration into tissue thus promoting cervical
remodeling and parturition by estrogen [58] Further studyis needed in terms of the effect of BSS on inflammatoryreactions and parturition
Furthermore theCG-DGherb pair has other names suchas Gunggui-tang (weight ratios of 2 3 or 1 1) Ogeum-san(1 1) Iphyo-san (1 1) and Sinmyo Bulsu-san (1 2) thoseare prepared at different weight ratios [3] Accordinglyweight ratio should be determined based on considerationsof targeted symptoms for relevant clinical applications
5 Conclusion
This study results show that Bulsu-san (BSS) is highlyconnected to the parturition related pathways biologicalprocesses and organsMost compounds inBSSwork togetherwith multiple target genes in a synergetic way and this wasconfirmed using herb-compound-target network and target-pathway network analysis The mRNA expression of relevanttarget genes of BSS was elevated significantly in parturition
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
Evidence-Based Complementary and Alternative Medicine 13
ADORA2A
HTR2A
CTSD
PIK3CG
PKM2
PTGS1
KCNMA1
PRKACA
PON1
GCGR
SERPINE1PRKCB
LCT
GLRA1
SmoothMuscleNR1I3
SLC6A3ESR1
NOS2
Pituitary
IL6
MOGAT2
WholeBlood
PCYT1APPARD
IL10
LTA4H
PPARA
MAPK14
PTGS2
RXRANQO2
CDC25B ADRB2
GSK3B
CAT
UCP2HMGCR
NCOA1
NR3C1ACSL4
ENPP7 CHRNA2
CHRM2ADSS
SRC
ERBB2ESR2
CHRNA7
hypothalamus andor pituitary
LEPREL2
NOS1
NCOA2
fetus andor placenta
SDHA
SP1
F7SLC6A4
MAT2A
HP PLG
GRIA2
SLC22A5
GABRA4
BCL2NOS3
ABAT
ME2
DRD1
HK1
NTRK2
CETP
LEPRE1
CASP3
GIG18
AKR1B1
TXNRD1
PTEN
FOSL2
SLC6A2
SCN5A
ADRA1A
AGXT
SUCLA2
RAC1
ME3
OAT
UCP3
MAOB KDR
ADH1B
EDNRA
CDK2
COL7A1
NR3C2
DPP4
ADRA1D
ADRB1
COL1A1SOAT1ADRA1B
CHEK1
CA1
CHAT
PIM1ADRA2A
PTGER3
AR
GABRA3SELP
CHI3L1P4HA2
PDE3A
PGR
ADRA2B
ATP5B
DRD2PDX1
TEP1
ENPEP
uterus andor uterus corpus
IGHG1
KCNH2
FOS
CITED1
CCNA2
F2
SLC2A2
PTPN1
MAOA
LPL
ADH1A
OTC
MAT1A
APOB
PNP
PPARG
CHRM3
GABRA1
BCHE
ALDH5A1
AHCYL1
IGF2
PDHX
ADRA2C
SREBF2
ALAD
PPARGC1A
GOT2
SOD1
SCDGLUD1
GAP43
GOT1
PKIA SREBF1HSP90AB1
GFAP
FABP1
G6PC
ACSL1
GABRA5
PLAU GABRA2
CALM1
Figure 5 Target organ location map it shows that tissue-specific patterns of mRNA expression are highly active in relative organs ofparturition process such as uterus fetus placenta hypothalamus pituitary and smooth muscle Yellow nodes show compounds and pinknodes indicate targets and node size is relative to the degree and edges demonstrate interactions between nodes
related organ tissues such as those of the uterus placentafetus hypothalamus and pituitary gland
This study employed the network analytical methodsto show the multicompound multitarget properties of BSSThe results not only support clinical applications of BSS oneasing childbirth but also suggest the related target genes andpathways of BSS on promoting parturition according to asystems-level in silico analytic approach However detailedmechanisms and other functions of BSS should be discussedfurther
Conflicts of Interest
The authors declare that there are no conflicts of interest
Acknowledgments
This research was supported by Basic Science ResearchProgram through the National Research Foundation ofKorea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01059994)
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
14 Evidence-Based Complementary and Alternative Medicine
References
[1] S Kim and Y Lee ldquoThe prescriptions of enriching blood andnourishing vital essence (Fill yin blood prescription)rdquo Journalof Korean Medical Classics vol 20 pp 67ndash77 2007
[2] Y Jin C Qu Y Tang et al ldquoHerb pairs containing AngelicaeSinensis Radix (Danggui) a review of bio-active constituentsand compatibility effectsrdquo Journal of Ethnopharmacology vol181 pp 158ndash171 2016
[3] J Lyu andC Jeong ldquoConstitution of prescription andmedicinaleffect adaptation diseases of rsquobullsoosan (berghean)rsquoin koreanmedical booksrdquo Journal of Korean Medical classics vol 29 pp17ndash41 2016
[4] W Li Y Tang J Guo et al ldquoEnriching blood effect comparisonin three kinds of blood deficiency model after oral administra-tion of drug pair of angelicae sinensis radix and chuanxiongrhizoma and each single herbrdquoChina Journal of ChineseMateriaMedica vol 36 no 13 pp 1808ndash1814 2011
[5] W Li M Huang Y Tang J Guo E Shang and X Liu ldquoEstab-lishment and optimization of acute blood stasis rat modelrdquoChinese Pharmacological Bulletin vol 12 p 032 2011
[6] Y-Z Hou G-R Zhao Y-J Yuan G-G Zhu and R HiltunenldquoInhibition of rat vascular smooth muscle cell proliferation byextract of Ligusticum chuanxiong andAngelica sinensisrdquo Journalof Ethnopharmacology vol 100 no 1-2 pp 140ndash144 2005
[7] C W C Bi L Xu X Y Tian et al ldquoFo Shou San an ancientchinese herbal decoction protects endothelial function throughincreasing endothelial nitric oxide synthase activityrdquoPLoSONEvol 7 no 12 Article ID e51670 2012
[8] W Li Y Hua Y Tang H Wang L Qian and M Gao ldquoEffectsof radix angelicae sinensis and rhizoma chuanxiong on mouseuterine contractions in vitrordquo Journal of Nanjing University ofTraditional Chinese Medicine vol 2 p 014 2010
[9] M McLean and R Smith ldquoCorticotrophin-releasing hormoneand human parturitionrdquo Reproduction vol 121 no 4 pp 493ndash501 2001
[10] S K Kota K Gayatri S Jammula et al ldquoEndocrinology of par-turitionrdquo Indian Journal of Endocrinology and Metabolism vol17 no 1 pp 50ndash59 2013
[11] M Ledingham A J Thomson I A Greer and J E NormanldquoNitric oxide in parturitionrdquo BJOG An International Journal ofObstetrics amp Gynaecology vol 107 pp 581ndash593 2000
[12] M Shen S Tian Y Li et al ldquoDrug-likeness analysis of tra-ditional Chinese medicines 1 property distributions of drug-like compounds non-drug-like compounds and natural com-pounds from traditional Chinese medicinesrdquo Journal of Chem-informatics vol 4 no 1 article 31 2012
[13] X XuW Zhang CHuang et al ldquoA novel chemometricmethodfor the prediction of human oral bioavailabilityrdquo InternationalJournal ofMolecular Sciences vol 13 no 6 pp 6964ndash6982 2012
[14] I Hubatsch E G E Ragnarsson and P Artursson ldquoDetermi-nation of drug permeability and prediction of drug absorptionin Caco-2 monolayersrdquo Nature Protocols vol 2 no 9 pp 2111ndash2119 2007
[15] W Tao X Xu X Wang et al ldquoNetwork pharmacology-basedprediction of the active ingredients and potential targets ofChinese herbal Radix Curcumae formula for application tocardiovascular diseaserdquo Journal of Ethnopharmacology vol 145no 1 pp 1ndash10 2013
[16] J Zhang Y Li S-S Chen et al ldquoSystems pharmacology dis-section of the anti-inflammatory mechanism for the medicinal
herb Folium eriobotryaerdquo International Journal of MolecularSciences vol 16 no 2 pp 2913ndash2941 2015
[17] PWillett JM Barnard andGMDowns ldquoChemical similaritysearchingrdquo Journal of Chemical Information and ComputerSciences vol 38 no 6 pp 983ndash996 1998
[18] H Liu J Wang W Zhou Y Wang and L Yang ldquoSystemsapproaches and polypharmacology for drug discovery fromherbal medicines an example using licoricerdquo Journal of Ethno-pharmacology vol 146 no 3 pp 773ndash793 2013
[19] K S Pang ldquoModeling of intestinal drug absorption roles oftransporters and metabolic enzymes (for the gillette reviewseries)rdquoDrugMetabolismampDisposition vol 31 no 12 pp 1507ndash1519 2003
[20] T Pei C Zheng C Huang et al ldquoSystematic understandingthe mechanisms of vitiligo pathogenesis and its treatment byQubaibabuqi formulardquo Journal of Ethnopharmacology vol 190pp 272ndash287 2016
[21] C A Lipinski F Lambardo B W Dominy and P J FeeneyldquoExperimental and computational approaches to estimate sol-ubility and permeability in drug discovery and developmentsettingsrdquo Advanced Drug Delivery Reviews vol 64 pp 4ndash172012
[22] A K Ghose V N Viswanadhan and J J Wendoloski ldquoPredic-tion of hydrophobic (lipophilic) properties of small organicmolecules using fragmental methods an analysis of ALOGPand CLOGPmethodsrdquoThe Journal of Physical Chemistry A vol102 no 21 pp 3762ndash3772 1998
[23] P Rajasethupathy S J Vayttaden and U S Bhalla ldquoSystemsmodeling a pathway to drug discoveryrdquo Current Opinion inChemical Biology vol 9 no 4 pp 400ndash406 2005
[24] H Yu J Chen X Xu et al ldquoA systematic prediction of multipledrug-target interactions from chemical genomic and phar-macological datardquo PLoS ONE vol 7 no 5 Article ID e376082012
[25] C H Wu R Apweiler A Bairoch D A Natale W C BarkerB Boeckmann et al ldquoThe universal protein resource (UniProt)an expanding universe of protein informationrdquo Nucleic AcidsResearch vol 34 pp D187ndashD191 2006
[26] G Bindea B Mlecnik H Hackl et al ldquoClueGO a Cytoscapeplug-in to decipher functionally grouped gene ontology andpathway annotation networksrdquoBioinformatics vol 25 no 8 pp1091ndash1093 2009
[27] M E Smoot K Ono J Ruscheinski P L Wang and T IdekerldquoCytoscape 28 new features for data integration and networkvisualizationrdquo Bioinformatics vol 27 no 3 pp 431-432 2011
[28] J-B Pan S-C Hu D Shi et al ldquoPaGenBase a pattern genedatabase for the global and dynamic understanding of genefunctionrdquo PLoS ONE vol 8 no 12 Article ID e80747 2013
[29] W Zhang Q Tao Z Guo et al ldquoSystems pharmacologydissection of the integrated treatment for cardiovascular andgastrointestinal disorders by traditional chinese medicinerdquoScientific Reports vol 6 p 32400 2016
[30] Z Prevorsek G Gorjanc B Paigen and S Horvat ldquoCongenicand bioinformatics analyses resolved a major-effect Fob3b QTLon mouse Chr 15 into two closely linked locirdquo MammalianGenome vol 21 no 3-4 pp 172ndash185 2010
[31] J Du B Bai X Kuang et al ldquoLigustilide inhibits spontaneousand agonists- or K+ depolarization-induced contraction of ratuterusrdquo Journal of Ethnopharmacology vol 108 no 1 pp 54ndash58 2006
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007
Evidence-Based Complementary and Alternative Medicine 15
[32] K J Chen and K Chen ldquoIschemic stroke treated with ligus-ticum chuanxiongrdquo Chinese Medical Journal vol 105 pp 870ndash873 1992
[33] W Li J Guo Y Tang et al ldquoPharmacokinetic comparison of fer-ulic acid in normal and blood deficiency rats after oral admin-istration of angelica sinensis ligusticum chuanxiong and theircombinationrdquo International Journal of Molecular Sciences vol13 no 3 pp 3583ndash3597 2012
[34] W Karl W Cathy X Li and G Amy ldquoPharmacokineticanalyses of ferulic acid in rat plasma by liquid chromatog-raphytandem mass spectrometry a synergistic action of anancient herbal decoction fo shou sanrdquo Pharmaceutica AnalyticaActa vol 6 no 5 p 2 2015
[35] D Huang ldquo8 purgative herbsrdquo Chinese Materia Medica Chem-istry Pharmacology and Applications p 231 1998
[36] X Liu Q Zhao C Li R Zhang X Wang and W XuldquoSynthesis of chuanxiong perlolyrine and its protective effecton injured vascular endothelial cellsrdquo Journal of ShandongUniversity (Health Sciences) vol 41 pp 485ndash487 2003
[37] J Ishida M Kozuka H-K Wang et al ldquoAntitumor-promotingeffects of cyclic diarylheptanoids on Epstein-Barr virus activa-tion and two-stage mouse skin carcinogenesisrdquo Cancer Lettersvol 159 no 2 pp 135ndash140 2000
[38] J Liu T Pei and J Mu ldquoSystems pharmacology uncoversthe multiple mechanisms of Xijiao Dihuang decoction for thetreatment of viral hemorrhagic feverrdquo Evidence-Based Comple-mentary andAlternativeMedicine vol 2016Article ID902503617 pages 2016
[39] H Wang and Y V Stjernholm ldquoPlasma membrane receptormediated MAPK signaling pathways are activated in humanuterine cervix at parturitionrdquo Reproductive Biology and Endo-crinology vol 5 article 3 p 1 2007
[40] SMesiano andTNWelsh ldquoSteroid hormone control ofmyom-etrial contractility and parturitionrdquo Seminars in Cell amp Develop-mental Biology vol 18 no 3 pp 321ndash331 2007
[41] PWNathanielsz S L Jenkins J D Tame J AWinter S Gullerand D A Giussani ldquoLocal paracrine effects of estradiol arecentral to parturition in the rhesus monkeyrdquo Nature Medicinevol 4 no 4 pp 456ndash459 1998
[42] Y Li H Je S Malek and K G Morgan ldquoERK12-mediatedphosphorylation of myometrial caldesmon during pregnancyand laborrdquoAmerican Journal of PhysiologymdashRegulatory Integra-tive and Comparative Physiology vol 284 no 1 pp R192ndashR1992003
[43] Y Li H-D Je S Malek and K G Morgan ldquoRole of ERK12in uterine contractility and preterm labor in ratsrdquo AmericanJournal of Physiology-Regulatory Integrative and ComparativePhysiology vol 287 no 2 pp R328ndashR335 2004
[44] S I Berger and R Iyengar ldquoNetwork analyses in systems phar-macologyrdquo Bioinformatics vol 25 no 19 pp 2466ndash2472 2009
[45] P Li L-W Qi E-H Liu J-L Zhou and X-D Wen ldquoAnalysisof Chinese herbal medicines with holistic approaches and inte-grated evaluationmodelsrdquoTrACTrends inAnalytical Chemistryvol 27 no 1 pp 66ndash77 2008
[46] L M Espinoza-Fonseca ldquoThe benefits of the multi-targetapproach in drug design and discoveryrdquo Bioorganic ampMedicinalChemistry vol 14 no 4 pp 896-897 2006
[47] W Zhang Y Bai Y Wang andW Xiao ldquoPolypharmacology indrug discovery a review from systems pharmacology perspec-tiverdquo Current Pharmaceutical Design vol 22 no 21 pp 3171ndash3181 2016
[48] S Wray K Jones and S Kupittayanant ldquoCalcium signalingand uterine contractilityrdquo Journal of the Society for GynecologicInvestigation vol 10 no 5 pp 252ndash264 2003
[49] G Y Oudit and Z Kassiri ldquoRole of PI3 kinase gamma inexcitation-contraction coupling and heart diseaserdquo Cardiovas-cular and Hematological Disorders vol 7 no 4 pp 295ndash3042007
[50] R Smith ldquoParturitionrdquo The New England Journal of Medicinevol 356 no 3 pp 271ndash283 2007
[51] S Y Suh and W G An ldquoSystems pharmacological approachof pulsatillae radix on treating crohnrsquos diseaserdquo Evidence-BasedComplementary and Alternative Medicine vol 2017 Article ID4198035 21 pages 2017
[52] C l Chen and J Y Chen ldquoAnalysis on the effect of foshousancombined with ethacridine lactate in mid-term pregnancyrdquoJournal of Practical Traditional ChineseMedicine vol 30 pp 511-512 2014
[53] Y Sun ldquoModified Xiong Gui Tang with auricular acupoint pressfor 70 cases of late pregnancyrdquo Guide of China Medicine vol 7no 20 pp 92-93 2009
[54] J-J Koksma J-M Fritschy V MacK R E Van Kesterenand A B Brussaard ldquoDifferential GABA A receptor clusteringdetermines GABA synapse plasticity in rat oxytocin neuronsaround parturition and the onset of lactationrdquo Molecular andCellular Neuroscience vol 28 no 1 pp 128ndash140 2005
[55] HMaulM LongoG R Saade andR EGarfield ldquoNitric oxideand its role during pregnancy From ovulation to deliveryrdquoCurrent Pharmaceutical Design vol 9 no 5 pp 359ndash380 2003
[56] M-J Ahn M K Lee Y C Kim and S H Sung ldquoThe simul-taneous determination of coumarins in Angelica gigas root byhigh performance liquid chromatography-diode array detectorcoupled with electrospray ionizationmass spectrometryrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 46 no 2 pp258ndash266 2008
[57] S Kim H Oh J Kim J Hong and S Cho ldquoA review of phar-macological effects of angelica gigas angelica sinensis angelicaacutiloba and their bioactive compoundsrdquo The Journal ofKorean Oriental Medicine vol 32 pp 1ndash24 2011
[58] J E Norman S Bollapragada M Yuan and S M NelsonldquoInflammatory pathways in the mechanism of parturitionrdquoBMC Pregnancy and Childbirth vol 7 supplement 1 article S72007