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Journal of Pharmacological Sciences 133 (2017) 115e121
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Journal of Pharmacological Sciences
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Full paper
Aconitine: A potential novel treatment for systemic
lupuserythematosus
Xiaodong Li a, b, 1, Liwei Gu a, 1, Lan Yang a, Dong Zhang a,
Jianying Shen a, *
a Institute of Chinese Materia Medica, China Academy of Chinese
Medical Sciences, Beijing 100700, Chinab Gansu Provincial Hospital
of Traditional Chinese Medicine, Lanzhou 730050, China
a r t i c l e i n f o
Article history:Received 6 September 2016Received in revised
form30 December 2016Accepted 21 January 2017Available online 24
February 2017
Keywords:AconitineSystemic lupus erythematosus
(SLE)Pristane-induced lupus (PIL)AutoantibodiesTraditional Chinese
medicine (TCM)
* Corresponding author. No. 16 Nanxiaojie, DongzhiChina.
E-mail address: [email protected] (J. Shen).Peer review under
responsibility of Japanese Pha
1 Equal contribution.
http://dx.doi.org/10.1016/j.jphs.2017.01.0071347-8613/© 2017 The
Authors. Production and hostinlicense
(http://creativecommons.org/licenses/by-nc-n
a b s t r a c t
Background: Aconitum plants have been widely used in China for
thousands of years. Recent evidencesindicate that aconitine, the
main active ingredient of Aconitum, has immunomodulatory properties
thatmight be useful for treating autoimmune diseases, such as
rheumatoid arthritis. In this study, we con-ducted a pilot study to
explore the effect and mechanisms of aconitine on the treatment of
systemiclupus erythematosus.Methods: A pristane-induced murine
model was used. The pristane-induced mice were treated
withaconitine (25, 75 mg kg�1 d�1, po) for 9 weeks. Every three
weeks, proteinuria was detected to monitorthe kidney damage and
blood was collected to measure serum levels of autoantibodies,
besides thekidney pathological examination. The major B cell
activating factor and major pro-inflammatory medi-ators, PGE2,
IL-17a and IL-6, were also detected.Results: We found that
aconitine significantly improved the mouse health, decreased the
elevated bloodleukocyte counts, reduced the serum level of
anti-double-stranded DNA (anti-dsDNA) antibody, greatlyameliorated
renal histopathologic damage and reduced IgG deposit in glomerular.
Furtherly, the levels ofPGE2, IL-17a and IL-6, were found to have
decreased in aconitine treated mice.Conclusion: We have
demonstrated that aconitine can inhibit the progression of disease
and amelioratethe pathologic lesion of systemic lupus
erythematosus.
© 2017 The Authors. Production and hosting by Elsevier B.V. on
behalf of Japanese PharmacologicalSociety. This is an open access
article under the CC BY-NC-ND license
(http://creativecommons.org/
licenses/by-nc-nd/4.0/).
1. Introduction
Systemic lupus erythematosus (SLE) is a chronic
autoimmunedisorder affecting multiple organs and characterized by a
variety ofautoantibodies, immune complex deposition in tissues and
subse-quent development of glomerulonephritis (1). Several
animalmodels, genetically engineered or chemical-induced, are
widelyused for SLE studies. Pristane (2, 4, 10,
14-tetramethylpentadecane)administration in Balb/c mice, known as
the environmentallyinduced model, mimics human idiopathic lupus
syndrome and ischaracterized by lupus-specific autoantibody
production alongwith arthritis, hemorrhagic pulmonary capillaritis,
proteinuria and
men Nei Ave., Beijing 100700,
rmacological Society.
g by Elsevier B.V. on behalf of Japad/4.0/).
glomerulonephritis (2,3). In addition, the autoantibody level
ofpristane-induced lupus in BALB/c is comparable to that found
inMRL/lpr mice, such as antiribonucleoprotein (RNP)
antibodies(anti-Su, anti-Sm, and anti-U1RNP), anti-double-stranded
(ds)DNA, anti-single-stranded (ss) DNA, and anti-histone (4,5).
Thismodel has been widely used for exploring the pathogenesis of
theSLE, which is still poorly understood.
Standard clinical therapies for SLE are glucocorticoids
combinedwith immunosuppressive agents, antimalarial drugs and
non-steroidal anti-inflammatory drugs. They often lead
immunotol-erance or exhibit various side-effects and disease
relapses aftertherapy discontinuation or tapered doses (6).
Belimumab (Ben-lysta®) is the only drug approved by the US Food and
DrugAdministration (FDA) for 56 years. It's a fully humanized
mono-clonal antibody that inhibits B cell activating factor, and
used fortreatment of autoantibody-positive SLE in adults (7). But
the price isunacceptable for most SLE patients. To search less
costly and moreeffective novel treatments, we turned to traditional
Chinese med-icine (TCM).
nese Pharmacological Society. This is an open access article
under the CC BY-NC-ND
http://creativecommons.org/licenses/by-nc-nd/4.0/http://creativecommons.org/licenses/by-nc-nd/4.0/mailto:[email protected]://crossmark.crossref.org/dialog/?doi=10.1016/j.jphs.2017.01.007&domain=pdfwww.sciencedirect.com/science/journal/13478613http://www.elsevier.com/locate/jphshttp://dx.doi.org/10.1016/j.jphs.2017.01.007http://creativecommons.org/licenses/by-nc-nd/4.0/http://dx.doi.org/10.1016/j.jphs.2017.01.007http://dx.doi.org/10.1016/j.jphs.2017.01.007
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X. Li et al. / Journal of Pharmacological Sciences 133 (2017)
115e121116
Aconitum plants (Ranunculaceae family) have been widely usedto
treat various diseases, such as rheumatism, knee pain,
wheezing,cough, cyanosis, chronic diarrhea, impotence, dense tinea,
herpeszoster, scabies and other disorders in China for thousands of
years(8). In TCM, the pharmacological effects of Aconitum plants
includereviving Yang for resuscitation, dispelling Wind to
eliminatedampness, warming Channels to expel coldness. Aconitine
(AC,MW: 645.74, molecular formation: C34H47NO11, chemical
structure:shown in Fig. 1A) is the main active component in
Aconitum plants(9). It has been used for treatment of pain and
inflammation (10). Inthis study, using pristane-induced murine
model, we have studiedthe therapeutic effects of aconitine in lupus
symptoms and ourresults for the first time have demonstrated that
aconitine may be apotential novel treatment for SLE.
2. Materials and methods
2.1. Materials
Aconitine (C34H47NO11, MW, 645.74) was supplied by the Na-tional
Institutes for Food and Drug Control (NIFDC), with a purity
of99.5%. Prednisone acetate tablets were purchased from
Topfondpharmaceutical (Zhumadian, Henan, China). Concanavalin A
(ConA) was supplied by Solarbio (Beijing, China). The Prostaglandin
E2(PGE2) levels were measured using an ELISA kit (Fanke
industrialco., LTD, Shanghai, China). Other chemicals and reagents
were ofanalytical grade.
2.2. Mice
Female BALB/c mice (aged 4e6 weeks) were purchased from theVital
River experimental animal technical co. (Beijing, China). Themice
were housed under specific pathogen-free conditions and fedstandard
rodent chow and water ad libitum. Mice were
injectedintraperitoneally (ip) with either 0.5ml of pristane
(Sigma, St. Louis,Missouri, USA) or saline (controls) and monitor
proteinuria every 4weeks to track the disease progression. Animal
experiments wereconducted according to the institutional ethical
guidelines for an-imal experiments and approved by the
Institutional Animal Care
Fig. 1. A, Chemical structure of aconitine. B, The difference of
body weight during the treatmgroup. C, Time course of clinical
characteristics and levels of proteinuria in BALB/c mice.
and Use Committee (IACUC) at Institute of Chinese Materia
Medica,China Academy of Chinese Medical Sciences.
2.3. Preparation of aconitine and treatment of animals
When proteinuria appeared obviously in disease model mice,mice
(n ¼ 12 per group) were treated with aconitine 75 mg/kg or25 mg/kg
body weight by oral gavage daily for nine weeks inexperimental
groups and the positive control group were admin-istered only with
prednisone acetate 6.3 mg/kg. The normal controland model control
mice (n ¼ 14) received the same volume of 0.5%sodium
carboxymethylcellulose.
2.4. Scoring of proteinuria
The assessment was performed every three weeks starting atthree
weeks after injection of pristane (or saline in normal
controlgroup). Proteinuria test stripes were supplied by Pearl
Riverchemical reagent Co. (Guangzhou, China). Urine was collected
bycatching stress and assessed by using semiquantitative scores
(for0 ¼ �, 1 ¼ ±, 2 ¼ þ, 3 ¼ þþ, 4 ¼ þþþ). The same standard
wasadopted in each test.
2.5. Detection of auto-antibodies
Serum anti-double-stranded DNA antibody was measured
withenzyme-linked immunosorbent assay (ELISA) (Cusabio Biotech
Co.Ltd, Wuhan, China). Results are presented as ng/ml employing
thestandard curve provided by the manufacturer.
2.6. Routine blood tests
For laboratory measurements, 100 ml of mouse whole blood
wascollected into tubes containing EDTA anticoagulant. Routine
bloodtests were immediately performed using a SYSMEX-XS800i
auto-mated hematology analyzer (Sysmex Corp., Hyogo, Japan).
ent. *, p < 0.05, **, p < 0.01, calculated versus Control
group; ##, p < 0.01, versus model
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X. Li et al. / Journal of Pharmacological Sciences 133 (2017)
115e121 117
2.7. Flow cytometry
Heparin anticoagulant blood was stained with
fluorochrome-conjugated monoclonal antibodies against surface
markers: APC-B220, PerCP-CD3e and PE-CD8a (eBioscience, San Diego,
Califor-nia, USA), FITC-CD4 and appropriate isotypic antibodies (BD
Phar-mingen, New Jersey, USA) were used as controls. After remove
thered blood cells, the centrifugal pellet was washed with
phosphatebuffer saline, and then resuspended in 0.3 ml of PBS
contain 0.02%FBS. The cell surface markers were analyzed using a
flow cytometer(Accuri C6, BD, New Jersey, USA).
2.8. Lymphoproliferation
The spleens of the mice were harvested after nine weeks
oftreatment. The spleens were mashed and passed through a 200-mesh
sterile sieve to prepare a single cell suspension. The cellswere
suspended in RPMI-1640 (Invitrogen, Paisley, Scotland, UK)medium
with 10% fetal bovine serum (FBS) and centrifuged. Thepelleted
cells were resuspended in Red Blood Cell Lysis BufferLysing Buffer
to lyse erythrocytes. After being washed in PhosphateBuffer
Solution (PBS), the total number of cells was calculated and2 � 106
cells were seeded in 96-well plates, then stimulated withConA (5
mg/ml) for 68 h. Lymphoproliferation was determined byMTT
assay.
The culture supernatant were collected after 68 h and keptat �80
�C for detecting PGE2 and cytokines.
2.9. Cytokine detection
Interleukin-6 (IL-6), IL-4, IL-10, IFN-g, TNF-a, IL-17a and
IL-2cytokines was detected using a mouse Th1/Th2/Th17
cytometricbead array kit (CBA; BD Biosciences, San Diego,
California, USA) andwas analyzed on a FACSCalibur flow cytometer.
Standard curveswere determined for each cytokine from a range of
10e5000 pg/ml.
2.10. Histopathology and immunohistochemistry assay
Kidney and spleen tissue samples were collected at the time
ofharvest and fixed in 10% buffered formalin, embedded in
paraffin.Hematoxylin and eosin (H&E), periodic acid-Schiff
stain (PAS) andMasson stain were to used check and define the
pathological stageof fibrosis of the specimen tissue.
For immunohistochemical staining, the kidney tissues (3 mmthick)
were stained with fluorescein-conjugated anti-mouse IgGand IgM
(Abcam, Cambridge, UK). To observe the distribution of
Fig. 2. Auto-ds-DNA levels in BALB/c mice. *p < 0.05, **p
< 0.01, calculate
collagen under and immunoglobulins deposition used a
lightmicroscope.
2.11. Statistical analysis
All group results are expressed as mean ± standard
deviation(SD), if not stated otherwise. ANOVA, Student's t test or
Fisher'sexact test (two tailed) were used for the comparison of
groupvalues. For comparing group values that did not follow
Gaussiandistribution, ManneWhitney's test (two tailed) was
used.
3. Results
3.1. Gross observation and body weight measurement
In the positive control group, prednisone induced
deleteriouseffects in this experiment, and two mice died at the end
of treat-ment. During the treatment, mice manifested emaciation,
listless-ness and dry and dull hair in model and prednisone
groups.However, aconitine-treated mice demonstrated glossy hair
(datanot shown) and less body weight loss (Fig. 1B).
3.2. Time course of proteinuria
Proteinuria levels were measured for detecting renal
injurydegree. The proteinuria levels in aconitine groups were
decreasedmore significantly than control group and prednisone
group. Aco-nitine has displayed positive effects after three weeks
administra-tion (p < 0.05) (Fig. 1C).
3.3. Time course of autoantibodies in BALB/c PIL
Serum levels of anti-dsDNA, anti-nuclear antibody (ANA)
andproliferating cell nuclear antigen (PCNA) antibodies were
measuredfor detecting autoantibodies production. It is found that
aconitinestrongly reduced the production of anti-dsDNA antibodies
in thesera of lupus mice after administration for three to six
weeks(Fig. 2). In this study, the used dosages of aconitinewere
nontoxic tomice. Because the weight changed in aconitine group is
less thancontrol group and all of aconitine group mice in good
shape bothphysically and psychologically during the experiment.
These resultsdemonstrated that aconitine significantly ameliorated
the symp-toms of SLE. However, ANA and PCNA levers were under
thedetection limits in our ELISA kits.
d versus control group; #, p < 0.05, ##, p < 0.01, versus
model group.
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X. Li et al. / Journal of Pharmacological Sciences 133 (2017)
115e121118
3.4. Effects of aconitine on blood cell subsets
This is an objective data from complete blood count as
per-formed on an automated instrument, including an automatedwhite
blood cell (WBC) differential count. It is found that aconi-tine
could significantly reduce the WBC count, neutrophil (NEUT)and
lymphocyte number (LYMPH). In addition, the average redblood cell
hemoglobin concentration (MCHC) also decreased inaconitine groups
(Supplementary Table 1). Prednison significantlyreduce the CD4 T
and B lymphocyte number, and aconitine has adirect inhibitory trend
in both lymphocyte subtypes by flowcytometry analysis (Table 1, and
FACS results not shown). Theseresults demonstrated aconitine could
suppress the immuneresponse mildly. Which would effect on staved
off SLE symptom inmice.
3.5. Effect of aconitine on PGE2 and cytokines production in
thecultured supernatant of splenocyte
To investigate further the role of PGE2 and Th1/Th2 cytokinesin
disease pathogenesis and the potential of aconitine as thera-peutic
agents in SLE, the effect of aconitine on inflammatoryfactor PGE2
production of splenocyte in aconitine treated micewere assayed. The
production of cytokine was measured at 68 hin ConA-stimulated
splenocytes. It was found that there was asignificant reduction of
inflammatory factor, particularly treatedmice with aconite (25
mg/kg) compared with model controls.Mixed culture supernatant in
each group was detected by FCM.IL-6, IL-2, IL-4, IL-10, IL-17a,
IFN-g and TNF-a were analyzed.After treatment with aconitine, IL-6
and IL-17a reduced signifi-cantly (Fig. 3).
Table 1The number of lymphocyte in blood.
Group Mice number Dose CD4-T (109
Control 14 e 2.56 ± 0.65Model 14 e 4.49 ± 1.61Prednison 10 6.3
mg/kg 2.77 ± 0.85Aconitine 12 25 mg/kg 3.43 ± 1.57
12 75 mg/kg 3.66 ± 2
The results shown are representative of all animals in each
group. Data represent the meaversus Model group.
Fig. 3. Effect of AC on the content of pro-inflammatory
mediators in the mixed cultured ssplenocytes were prepared and
co-cultured with ConA 5 mg/ml for 68 h, then the
cytokinessignificantly suppress the IL-6 and IL-17a level. Values
are the mean ± SD of animals in eacgroup; #, p < 0.05, ##, p
< 0.01, versus model group.
3.6. Effects of aconitine treatment on histopathology of spleen
andkidney
Tissues of spleen and kidney were collected at the end of
nine-week administration of aconitine. Histological
examinationexhibited by H&E, PAS and Masson staining (Fig. 4).
Pristane in-duces a lupus-like syndrome in nonautoimmune mice
whoseglomerular basementmembrane thickening withmesangial
matrixexpansion and mesangial cell proliferation. Moreover, the
thick-ening of the glomerular capillary basement membrane were
alsonoted. However, aconitine can completely correct these
seriouskidney injuries. PAS and Masson staining reached a
similarconclusion.
3.7. Aconitine reduced the production of lupus-associated IgG
butnot IgM autoantibodies
IgG accumulation were found within the kidney tissue in
pris-tane induced lupus mice. It was found two of ten mice in 6.3
mg/kgprednisone treated group. Unlike mice in model group (8/14),
therewere only few (3/12) obvious abnormal changes were
evidencedafter completion of aconitine administration (Fig. 5AeE).
Further-more, the deposition of IgM autoantibodies was not detected
inkidney in our experiment (data not shown). It is suggest that
aco-nitine could relieve immune globulin deposition in kidney.
4. Discussion
Systemic lupus erythematosus (SLE) is an autoimmune diseasewhich
is potentially fatal. Over the past several decades, tremen-dous
enthusiasm and efforts have been devoted to tackling the
/l) CD8-T (109/l) B cell (109/l) CD4/CD8
0.62 ± 0.19 1.5 ± 0.66 4.21 ± 0.61** 1.19 ± 0.46** 2.51 ± 0.9**
3.82 ± 0.57## 0.92 ± 0.29** 1.75 ± 0.57# 3.05 ± 0.53
0.9 ± 0.33* 1.92 ± 0.71 3.75 ± 0.680.92 ± 0.48* 1.93 ± 0.99 4 ±
0.71
n ± SD.*p < 0.05, **p < 0.01, calculated versus Control
group; #, p < 0.05, ##, p < 0.01,
upernatant of splenocytes from BALB/c mice in vitro. A. IL-6, B.
IL-17a, C. PGE2. Miceproduction were determined. Values are the
mixed of animals in each group. Aconitineh group, each performed in
triplicate. *, p < 0.05, **, p < 0.01, calculated versus
control
-
Fig. 4. Aconitine ameliorates kidney injury in pristane induced
mice. (A) Control, (H&E � 400). (B) Model, showing glomerular
basement membrane thickening with mesangialmatrix expansion and
mesangial cell proliferation. The glomerular capillary basement
membranes were thickening (H&E � 400). (C) 6.3 mg/kg prednisone
treated mice, beside theinjuries in model mice, protein cast even
appeared (H&E� 400). (D) 25 mg/kg aconitine (H&E� 400) and
(E) 75 mg/kg aconitine treated mice completely correct these
serious kidneyinjuries (H&E � 400). (F) Control, (PAS � 400).
(G) Model, glomerular basement membrane thickening with mesangial
matrix expansion and mesangial cell proliferation(PAS � 400). (H)
75 mg/kg aconitine treated group (PAS � 400). (I) Control, showing
normal kidney structure (Masson � 400). (J) Model (Masson � 400),
showing glomerularbasement membrane thickening with mesangial
matrix expansion and mesangial cell proliferation. Blue-staining
are increased collagen fibers. The glomerular capillary
basementmembranes were thickening. (K) 25 mg/kg aconitine (Masson �
400) and (L) 75 mg/kg aconitine treated mice completely correct
these serious kidney injuries. The pathologicalsections did not
show abnormal kidney structures (Masson � 400).
X. Li et al. / Journal of Pharmacological Sciences 133 (2017)
115e121 119
numerous challenges to understand the etiopathogenesis of
SLE.However, the precise mechanism of this disease is still
unclear.
In this pilot study, pristane induced mice displayed the
symp-toms of SLE. The IgG induced by pristane, targeted to a
variety ofnuclear components including dsDNA, ssDNA and
nucleoproteins.Anti-dsDNA antibodies are highly specific for lupus
and are impli-cated in the pathogenesis of lupus nephritis (11).
The prevalence ofanti-dsDNA antibody is 70e80% in SLE patients, so
we chose it forthe important indicator in our study. The anti-dsDNA
antibodies inthe sera of normal mice, measured before pristane
injection, and inthe sera of SLE model mice, measured before
treatment, were6.9 ± 1.77 ng/ml and 14.39 ± 4.44 ng/ml,
respectively (Data notshown), indicating the establishment of SLE
model induced bypristane.
In Traditional Chinese medicine, Aconitum plants have beenwidely
used to treat diseases such as rheumatism and knee pain.Aconitine,
which is useful for treatment of pain and inflammation(10), is the
main active component in Aconitum plants (9). However,the clinic
application of aconitine is limited by its high toxicity. Itslethal
dose 50% (LD50) for mice is 1.8 mg/kg (orally) and 0.308 mg/kg
(intraperitoneally) (12). In a long-term administration study
thatan oral dose of 1 mg/kg per day was administered to mice for
22days, the frequency of arrhythmias remarkably decreasedwith
timeand repeated administration of aconitine (13). To minimize
thetoxicity of aconitine, we used oral dose of 75 mg/kg or 25 mg/kg
bodyweight for 9 weeks. No death or toxicity was detected during
andafter the treatment. During the treatment, mice manifested
ema-ciation, listlessness and dry and dull hair in model and
prednisone
-
Fig. 5. Kidney accumulation of IgG. (A) Control, (B) Model, (C)
6.3 mg/kg prednisone treated mice, (D) 25 mg/kg aconitine and (E)
75 mg/kg aconitine treated mice. The positive cellsdisplayed
brownish yellow granules on the cell surface and/or cytoplasm
(original magnification, �400).
X. Li et al. / Journal of Pharmacological Sciences 133 (2017)
115e121120
groups. However, aconitine treatment ameliorated the
diseasesymptoms, and decreased anti-dsDNA levels (Fig. 2).
Aconitine-treated mice demonstrated glossy hair (data not shown)
and lessbody weight loss (Fig. 1B).
Unexpectedly, during the treatment, the control mice treatedwith
0.5% sodium carboxymethylcellucose were found to loss theweight
about 2 g. It may be caused by the age. As to Berlin FatMouse
Inbred line 860 (BFMI860) mice, they reach the growth peakat the
age of day 300 and begin to loss the body weight (14). In ourstudy,
the BABL/c mice were at the age of day 240 when wemeasured the body
weight last time, and they may have passed thegrowth peak and begin
to loss the body weight. Compared withmice in other three groups,
aconitine-treated mice demonstratedless body weight loss (Fig.
1B).
One of the most common clinical feature of SLE is lupusnephritis
(glomerulonephritis), which can be seen in up to 60% ofall SLE
patients and this complications require hemodialysis andcause
higher lethality (15). Lupus nephritis is thought to
involveglomerular inflammation induced by immune complexes
andcomplement deposition (16). This type of nephritis is diagnosed
bythe presence of biomarkers for kidney damage such as
proteinuria,creatinine and blood urea nitrogen. Recently, new
urinary bio-markers, such as the inflammatory mediators IL-6,
vascular celladhesion molecule-1 and urinary immune cells, have
been gradu-ally accepted (17,18).
Present study has shown that aconitine could
significantlysuppress the proliferation of glomerular mesangial
cells andmaintain the glomerular capillary basement membranes. It
hasgreatly ameliorated renal histopathologic damage and reduced
IgGdeposit in glomerular. Overall, these data suggest that
aconitinetreatment could inhibit the activity and progress of SLE
and therebyimprove their quality of life.
Many reports suggested that cytokines may act as key players
inthe immunopathogenesis of SLE (19,20). IL-17 is a potent
pro-inflammatory cytokine produced by activated T lymphocytes.
Itplays a critical role in recruit monocytes and neutrophils to the
siteof inflammation, inducing production of proinflammatory IL-6
(21),and driving B-cell differentiation into plasma cells and
productionof autoantibodies (22). IL-6 has a range of biological
activities on
various target cells that plays an important role in immune
regu-lation and inflammation. Data from several studies suggest
that IL-6 plays a critical role in the B cell hyperactivity and
immunopa-thology of human or murine models of SLE, induction of IgG
pro-duction (23), and may even have a direct role in mediating
tissuedamage (24). Meanwhile, IL-6 induces naïve CD4þ Tcells to
developinto Th17 cells, the IL-17-producing cells (25). Consistent
withprevious report, we have observed amarked increase in levels of
IL-6 in the pristane induced BALB/c mice (26). PGE2 is a bioactive
lipidwhich can elicit a wide range of biological effects,
includinginflammation, cell proliferation, apoptosis and
angiogenesis (27).Recent reports have elaborated PGE2 could promote
the activationof Th17 cells and production of IL-17 (28). In
accordance with manyother studies, we have demonstrated directly
that PGE2 couldenhance IL-17a expression (29,30). Aconitine could
inhibit IL-17aand IL-6 secretion, and thereby suppress B-cell
hyperactivity andautoantibodies production. However, there is
little influence inother Th1/Th2 cytokines, such as IL-2, TNF-a and
IFN-g. So wepropose the influence of PGE2, IL-17a and IL-6 may be
amechanismof aconitine action on SLE.
In conclusion, this pilot study for the first time has
demon-strated that aconitine is effective for the treatment of SLE.
Aco-nitine raised the body weight, improved health status,
decreasedthe number of blood leucocytes, decreased the serum level
ofanti-dsDNA antibody, ameliorated the renal pathology, andthereby
relieved bodies suffering damage. These effects of aco-nitine might
be achieved in part by decreasing the inflammationin kidney and
inhibiting B cells activation through suppressingthe major
pro-inflammation mediators, IL-17a, IL-6, and PGE2.Further studies
will be required to confirm the efficacy andmechanism of action of
aconitine, as to provide evidence for itspotential clinical
application for human systemic lupus erythe-matosus in the
future.
Author contribution
Conceived and designed the experiments: Xiaodong Li, Liwei Guand
Jianying Shen. Contributed reagents/materials/analysis tools:Lan
YANG. Performed the experiments: Xiaodong Li, Liwei Gu and
-
X. Li et al. / Journal of Pharmacological Sciences 133 (2017)
115e121 121
Dong Zhang. Analyzed the data: Jianying Shen, Liwei Gu
andXiaodong Li. Wrote the paper: Jianying Shen and Liwei Gu.
Conflict of interest
There is no conflict of interest.
Acknowledgments
This work was supported by National Science and TechnologyMajor
Projects for “Major NewDrugs Innovation and Development”(Project
No. 2013ZX09301307001002) and National Natural ScienceFoundation of
China (Grant No. 81402075) and China PostdoctoralScience Foundation
funded project (2014M561160). The authorsthank Lanfang Li, Guihua
Yu, Canghai Lee and Shuying Guo for theirtechnical help.
Appendix A. Supplementary data
Supplementary data related to this article can be found at
http://dx.doi.org/10.1016/j.jphs.2017.01.007.
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Aconitine: A potential novel treatment for systemic lupus
erythematosus1. Introduction2. Materials and methods2.1.
Materials2.2. Mice2.3. Preparation of aconitine and treatment of
animals2.4. Scoring of proteinuria2.5. Detection of
auto-antibodies2.6. Routine blood tests2.7. Flow cytometry2.8.
Lymphoproliferation2.9. Cytokine detection2.10. Histopathology and
immunohistochemistry assay2.11. Statistical analysis
3. Results3.1. Gross observation and body weight measurement3.2.
Time course of proteinuria3.3. Time course of autoantibodies in
BALB/c PIL3.4. Effects of aconitine on blood cell subsets3.5.
Effect of aconitine on PGE2 and cytokines production in the
cultured supernatant of splenocyte3.6. Effects of aconitine
treatment on histopathology of spleen and kidney3.7. Aconitine
reduced the production of lupus-associated IgG but not IgM
autoantibodies
4. DiscussionAuthor contributionConflict of
interestAcknowledgmentsAppendix A. Supplementary dataReferences