Rajiv Gandhi University of Health Sciences
Rajiv Gandhi University of Health Sciences
Karnataka, Bengaluru
“PHARMACOLOGICAL AND TOXICOLOGICAL STUDIES OF FRUITS OF LUFFA
ACUTANGULA.ROXB ON RATS”
A Protocol submitted to Rajiv Gandhi University of Health
Sciences Karnataka, Bengaluru
In partial fulfillment of the requirement for the award of
MASTER OF PHARMACY
IN
PHARMACOLOGY
KANAGARLA. N. P. GOWTHAM
Department of Pharmacology,
National College of Pharmacy,
Balraj-Urs Road,
Shimoga-577 201Karnataka-INDIA
Rajiv Gandhi University of Health Sciences, Karnataka,
Bengaluru
Annexure – II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
01
Name and Address of the Candidate
K. N. P. GOWTHAM
S/O M. V. B. P. KUTUMBA RAO
DOOR NO: 8-149,
MALLAYA PETA
RAJAHMUNDRY-533105
02
Name of the Institution
NATIONAL COLLEGE OF PHARMACY,
BALRAJ-URS ROAD,
SHIMOGA-577 201KARNATAKA-INDIA
03
Course of the Study
Branch
M. Pharmacy, (Pharmacology)
04
Date of Admission to course
04-10-2010
05
Title of the Topic
“PHARMACOLOGICAL AND TOXICOLOGICAL STUDIES OF FRUITS OF LUFFA
ACUTANGULA.ROXB ON RATS.”
06
Brief resume of the intended work
6.1. Need for the Study
Enclosure – I
6.2. Review of the Literature
Enclosure – II
6.3. Objective of the Study
Enclosure – III
07
Materials and Methods
7.1. Source of data
Enclosure – IV
7.2. Methods of collection of data
Enclosure – V
7.3. Does the study require any
Investigations on animals?
If yes give details
Enclosure – VI
7.4. Has ethical clearance been
obtained form your institution
in case of 7.3.
Enclosure – VI – A
08
List of References (About 4 – 6)
Enclosure – VII
09
Signature of the Candidate
10
Remarks of the Guide
The present research work is original and not published in any
of the journals. This work can be carried out in our Pharmacology
Department laboratory.
11
Name and Designation of
(in Block Letters)
11.1. Guide
11.2.Signature
11.3.Co-Guide (if any)
11.4.Signature
11.5. Head of the Department
11.6. Signature
Dr. I. J. KUPPAST
M.Pharm ,Ph.D , F.I.C.
PROFESSOR
National College of Pharmacy,
Balraj-Urs Road,
Shimoga-577 201
Karnataka-INDIA
N. A.
N. A.
Dr. K. L. MANKANI
M.Sc , Ph.D
Head of the Department
National College of Pharmacy,
Balraj-Urs Road,
Shimoga-577 201
Karnataka-INDIA
12
Remarks of the Principal
12.1. Signature
The present study is permitted to perform in the Pharmacology
Department laboratory of our Institution and the study protocol has
been approved by IAEC.
Principal
Enclosure – I
Brief resume of intended work:
6.1. Need for study:
Plants have had and still have a key role in history of life on
earth. Plants have been part of our lives since the beginning of
time; we get numerous products from plants, most of them not only
beneficial but also crucial to our existence1. Before the onset of
synthetic era, man was completely dependent on medicinal herbs for
prevention and treatment of diseases2. Plants have evolved the
ability to synthesize chemical compounds that help them, defend
against attack from a wide variety of predators such as insects,
fungi, herbivorous mammals. By chance, some of these compounds
whilst being toxic to plant predators turnout to have beneficial
effects when used to treat human diseases3. The use of plants to
heal or combat illness is as old as humankind. In the present
scenario, the demand for herbal products is growing throughout the
world and major pharmaceutical companies are currently conducting
extensive research on plant materials for their potential medicinal
value4.
Luffa acutangula.ROXB is believed to have originated in India,
where wild types still occur, but has now spread pantropically to
all areas with a high rainfall. It is cultivated and locally
naturalized in West Africa, from Sierra Leone to Nigeria. It is
cultivated from the coastal areas to the semi-dry savanna, e.g. in
Sierra Leone, Ghana, Benin and Nigeria.
Luffa acutangula.ROXB belongs to family Cucurbitaceae. Luffa
acutangula is a large monoious climber. It is one of the vegetable
plants with medicinal property. The stem of Luffa acutangula is 5
angled glabrous stem and trifid tendrils. Leaves are
orbicular-cordate, palmately 5-7 lobed, scabrous on both sides with
prominent veins and vein lets. The leaf juice is applied to skin
infections such as eczema. Leaf sap is used as an eye wash to cure
conjunctivitis, where as in some parts of West Africa leaf extract
is applied on sores caused by guinea worms to kill the parasite.
Flowers are yellow, males arranged in 12-20 flowered axilary
racemes. Female flowers are solitary, arranged in axils of males.
Ovary is strongly ribbed. Fruits are obovate, pale yellow brown in
color, 4-10 cm long and 2-4 cm broad and outer surface being
covered with 8-10 prominent longitudinal ribs. Fruit is divided
into 3 chambers; inner part is fibrous and easily detachable as a
whole from the outer part. Taste is bitter and transverse section
shows papillose epidermis with striated thick cuticle. Immature
fruits are used in cooking and also in soups and sauces. Young
fruits are eaten raw by cultivators and small fruits are sometimes
pickled. Seeds are black, ovoid-oblong, much compressed and winged.
Seeds yield an edible oil that is, however, sometimes bitter and
toxic. The fruits and seeds are sometimes used in herbal
preparation for the treatment of venereal diseases, particularly
gonorrhea. From the literature survey it is said to possess
ribosome inactivating protein [Luffin A and B] and terpenes with
immunomodulatory, anti-HIV, other pharmacological actions viz.
Antidiabetic, antioxidant, anticancer. Other uses like as an
ornamental climber for enclosure, mature dried fruits processed
into sponges and used for scrubbing the body or for domestic
purposes like washing utensils and as filters for local drinks such
as palm wine. Industrial use is made of these fibers for making
hats5-6.
The claim of anti-ulcer and hypolipidaemic activity and
toxicological studies of fruits has not been studied so far
scientifically. Hence the present study is aimed to investigate
anti-ulcer and hypolipidaemic activity and toxicological studies of
fruits of Luffa acutangula.ROXB.
Enclosure – II
6.2. Review of literature:
1. The head space volatile compounds of flowers of ridge gourd
and bitter gourds were analyzed by solid phase micro extraction
[SPME] coupled with Capillary Gas Chromatography/Mass Spectrometry
[GC-MS]. In ridge gourd 16 volatiles were identified and 9 were
tentatively identified which are of trans β-ocimene7.
2. Hepatoprotective activity of Luffa acutangula against CCl4
and Rifampicin induced liver toxicity in rats: a biochemical and
histopathological evaluation8.
3. Studies on the constituents of Luffa acutangula.ROXB.
Structures of acutosides A—G oleanane type triterpene saponins
isolated from herb9.
4. X-ray sequence and crystal structure of luffaculin-1, a novel
type 1 ribosome inactivating protein10.
5. Two proteins with ribosome inactivating, cytotoxic and
abortifacient activities from seeds of Luffa acutangula. ROEM
[Cucurbitaceae] 11.
6. Protein with abortifacient, ribosome inactivating, immuno-
modulatory, anti-tumor and anti-AIDS activities from Cucurbitaceae
plants12.
7. Apoptosis inducing activity of Luffa acutangula fruit in
leukemia cells13.
8. A Chitotetrose specific lectin from Luffa acutangula
Physico-chemical properties and the assignment of sugars in lectin
binding site14.
9. Isolation, macromolecular properties and combining site of a
chito-oligosaccharide specific lectin from exudates of ridge
gourd15.
10. Relationships of Aulacophora beetles feeding behavior with
Cucurbitaceae type in host crop16.
11. Luffangulin, a novel ribosome inactivating peptide from
ridge gourd seeds17.
12. Crystallization and preliminary X-ray analysis of
luffaculin, a ribosome inactivating protein from sponge gourd
seeds18.
13. Trypsin inhibitors from ridged gourd seeds: purification,
properties and amino acid sequence19.
14. Antimicrobial activity of whole plant of Luffa cylindrica
against some common pathogenic micro-organisms20.
15. Combining ability studies in ridge gourd21.
16. Comparative anti-diabetic activity of some herbal plant
extracts22.
17. Studies on the anti-microbial and anti-fungal activities of
the ethanolic extract of Luffa cylindrica fruit23.
18. Luffa acutangula.ROXB. Tea promotes developmental toxicity
to rats24.
19. PAF receptor binding antagonist activity of Malaysian
medicinal plant25.
20. Ethno medicines for jaundice used in tribal area of North
Maharashtra26.
21. Pharmacological potential of plants used in treatment of
piles27.
22. Antioxidant activity of 5 vegetables- traditionally consumed
by south Asian migrants in Bradford, Yorkshire, UK28.
23. Bilateral thalamic necrosis following ingestion of ridge
gourd infested with coelomycete fungi [diploid] 29.
Enclosure – III
6.3. Objectives of study:
1. Collection and authentication of Luffa acutangula.ROXB.
2. Drying of fruits of Luffa acutangula.ROXB.
3. Extraction of whole plant using different solvents.
4. Phytochemical investigations of extracts.
5. Screening of anti-ulcer and hypolipidaemic activity.
6. Evaluation of Toxicological studies.
Enclosure – IV
MATERIALS AND METHODS:
7.1. Source of data:
The required data will be obtained from:
1. Electronic data [internet].
2. Published Research Papers.
3. Review and Research Articles from Journal.
4. Library, National College of Pharmacy, Shimoga, Karnataka,
India.
Enclosure – V
7.2. Method of collection of data:
1. The plant of Luffa acutangula will be collected from local
areas of Shimoga district, Karnataka.
2. Screening of Pharmacological activities – i.e., anti-ulcer
and hypolipidaemic activity.
3. Evaluation of Toxicological studies.
4. Pharmacological studies include– hypolipidaemic activity and
anti-ulcer activity.
Hypolipidaemic activity will be carried out with obese rats; the
serum cholesterol level is examined after a period of time.
Activity will be concluded by comparing the results of control,
standard and test samples.
Anti-ulcer activity will be carried out either by pyloric
ligation or by drug induction method. Ulcer Index will be
calculated in the control and treated group.
Toxicological studies will be carried out as per the standard
procedure as per OECD and finally the results are compared with
treated and control group.
The results will be analyzed by ANOVA test.
Enclosure – VI
7.3. Does the study require any investigation or intervention to
be conducted on patients or other humans or animals?
As per the standard procedure, to study the pharmacological and
toxicological effects of fruits of Luffa acutangula will be carried
out on the Wistar albino rats.
7.4. Has ethical clearance been obtained from your
institution?
Ethical clearance is provided by the Institution.
Clearance number: NCP/IAEC/CL/22/05/2011-12.
Enclosure –VII
List of references:
1.
http://evolvingwellness.com/posts/984/the-value-and-importance-of-plants-in-medicine/
2. http://www.botanical-online.com/theimportanceofplants.htm
3. http://en.wikipedia.org/wiki/herbalism
4. Kokate C K. Text book of Pharmacognosy, Nirali Prakashan, New
Delhi. Ed.4. p. 3-4.
5.
http://database.prota.org/dbtw-wpd/exec/dbtwpub.dll?AC=QBE_QUERY&BU=http://database.prota.org/search.htm&TN=PROTAB~1&QB0=AND&QF0=Species+Code&QI0=Luffa+acutangula&RF=Webdisplay
6. Jyothi V, Srinath Ambati and Asha Jyothi V, “The
Pharmacognostic, Phytochem-ical and pharmacological profile of
Luffa acutangula”, International Journal of Pharmacy and
Technology; 2010, 2(4): 512-524.
7. Fernando L N and Grun I U, “Head space-SPME analysis of
volatiles of the ridge guard [Luffa acutangula] and bitter guard
flowers”, Flavour and Fragnance Journal; 2001, 16(4): 289-293.
8. Vishal B Jadhar, Vishnu N Thakare, Apuman A Suralkar, Avinash
D Deshpandae and Suresh R Naik, “Hepatoprotective activity against
CCl4 and Rifampicin induced liver toxicity in rats”, Indian Journal
of Experimental Biology; 2010, 48(8): 822-829.
9. Naga O T, Tanaka R, Iwase Y, Hanazono H and Okabe H, “Studies
on the constituents of Luffa acutangula.ROXB structures of
acutosides A—G oleanane type triterpene, saponins isolated from
herb”, Chemical and Pharmaceutical Bulletin(Tokyo); 1991, 39(3):
599-606.
10. Hou X, Chen M, Chen L, Meehan E J, Xie J and Haung M, “X-ray
sequence and crystal structure of luffaculin-1, a novel type 1
ribosome inactivating protein”, BMC structural biology; 2007, 7:
29.
11. Ng T B, Wong R N and Yeung H W, “Two proteins with ribosome
inactivating, cytotoxic and abortifacient activities from seeds of
Luffa acutangula.ROEM [Cucurbitaceae]”, Biochemistry International;
1992, 27(2): 197-207.
12. Ng T B, Chan W Y and Yeung H W, “Protein with abortifacient,
ribosome inactivating, immuno-modulatory, anti-tumor and anti-AIDS
activities from Cucurbitaceae plants”, General Pharmacology; 1992,
23(4): 597.
13. Purushotham Reddy B, Raghuram Reddy A, Srinivas Reddy B,
Venkata Mohan S and Sarma P N, “Apoptosis inducing activity of
Luffa acutangula fruit in leukemia cells”, International Journal of
Pharmaceutical Research and Development; 2010, 2(10): 109-122.
14. Vella Reddy Anatharam, Sankhavaram R Patanjali and Avadesha
Surolia, “A Chitotetrose specific lectin from Luffa acutangula
physico-chemical properties and the assignment of sugars in lectin
binding site”, Proc. Int. Symp. Biomol. Struct. Interactions,
Supplement Journal of Bioscience; 1985, 8(1,2): 403-411.
15. Vellareddy Anantharam, Sankhavaram R Patanjali, M Joginadha
Swamy, Ashik R Sanadi, Irwin J Goldstein and Avadesh Surolia,
“Isolation, macromolecular properties and combining site of a
chito-oligosaccharide specific lectin from exudates of ridge
gourd”, The Journal of Biological Chemistry; 1986, 261(31):
14621-14627.
16. Yang X, Kong C, Liang W, Zhang M and Hu F, “Relationships of
Aulacophora beetles feeding behavior with Cucurbitaceae type in
host crop”, Yengyongsheng Taixue Bao; 2005, 16(7): 1326-1329.
17. Wang H and Ng T B, “Luffangulin, a novel ribosome
inactivating peptide from ridge gourd seeds”, Life Science; 2002,
70(8): 899-906.
18. Ma Q, Yao G, Wus, LiH, LiJ and Dong Y, “Crystallization and
preliminary X-ray analysis of luffaculin, a ribosome inactivating
protein from sponge gourd seeds”, Acta Crystallogr D Biol
Crystallogr; 2000, 56(2): 185-186.
19. Haidar U C, Saha S K, Beavis R C and Sinha N K, “Trypsin
inhibitors from ridged gourd seeds: purification, properties and
amino acid sequence”, Journal of Protein Chemistry; 1996, 15(2):
177-184.
20. Indumathy R, Satheesh Kumar D, Kolagani Pallavi and
Sashikala Devi G, “Antimicrobial activity of whole plant of Luffa
cylindrica against some common pathogenic micro-organisms”,
International Journal of Pharmaceutical Sciences and Drug Research;
2011, 3(1): 29-31.
21. Rao Y N and Reddy P V, “Combining ability studies in ridge
gourd”, International Journal of Tropical Agriculture; 2000, 18(2):
141-146.
22. Patil Priyanka S, Patel M M and Bhavasar C J, “Comparative
anti-diabetic activity of some herbal plant extracts”, An
International Journal of Pharmaceutical Sciences; 2010, 1(1):
12-19.
23. Sashikala Devi G, Kottai Muthu A, Satheesh Kumar D, Rekha S,
Indumathy and Nandhini R, “Studies on the anti-microbial and
anti-fungal activities of the ethanolic extract of Luffa cylindrica
fruit”, International Journal of Drug Development and Research;
2009, 1(1):105-109.
24. Luciana Cristina Borges Fernandes, Luiz Augusto Vieira
Cordeiro and Benito Soto-Blanco, “Luffa acutangula.ROXB. Tea
promotes developmental toxicity to rats”, Journal of Animal and
Veterinary Advances; 2010, 9(8): 1255-1258.
25. Jantan I, Rafi I A and Jalil J, “PAF receptor binding
antagonist activity of Malaysian medicinal plant”, Phytomedicine:
International Journal of Phytotherapy and Phytopharmacology; 2005,
12(1, 2): 88-92.
26. Badgujarr S B and Patil M B, “Ethno medicines for jaundice
used in tribal area of North Maharashtra”, Natural Product
Radiance; 2008, 7(1): 79-81.
27. Lather A, Gupta V, Garg S, Singh A and Sachdeva K,
“Pharmacological potential of plants used in treatment of piles”,
Journal of Natura Conscientia; 2011, 2(1): 255-265.
28. Ansari N M, Houlihan L, Hussain B and Pieroni A,
“Antioxidant activity of 5 vegetables- traditionally consumed by
South Asian migrants in Bradford, Yorkshire, UK”, Phytotherapy
Research; 2005, 19: 907-911.
29. De Souza A, Naruencar K P, Fernandes Y and Arun G,
“Bilateral thalamic necrosis following ingestion of ridge gourd
infested with coelomycete fungi [diploid]”, Journal of Neurological
Science; 2010, 295(1,2): 104-106.