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DEPARTMENT OF PHARMACOLOGY
VYDEHI INSTITUTE OF MEDICAL SCIENCES & RESEARCH CENTRE
#82, EPIP Area, Nallurahalli, Whitefield, Bangalore - 560066
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BANGALORE, KARNATAKA.
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Name of the Candidate and
Address
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Name of the Institution
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Course of Study and Subject
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Date of Admission to
Course
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Title of the Topic
Dr. ANAND. M. INGALE.
DEPARTMENT OF PHARMACOLOGY,
VYDEHI INSTITUTE OF MEDICAL SCIENCES
AND RESEARCH CENTRE,
#82, EPIP AREA, NALLURAHALLI, WHITEFIELD,
BANGALORE-560066.
Vydehi Institute of Medical Sciences &
Research Centre, Bangalore.
M.D. PHARMACOLOGY
22-04-2011
Experimental evaluation of anti-ulcer activity of
Grape Seed Extract in Wistar Albino Rats.
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6. BRIEF RESUME OF THE INTENDED WORK:
6.1 NEED FOR THE STUDY:
Antioxidants are compounds that inhibit oxidation process and thereby eliminate the threat of
pathological processes. During environmental stress, elevation of corticosteroids accelerates
the generation of free radicals and suppresses immune function. Antioxidants are effective in
sheerful conditions, as they readily give up their own electron to free radicals. It has been
found that oxygen-derived free radicals are implicated in the mechanism of acute and chronic
gastric mucosal ulceration, and scavenging these free radicals plays an appreciable role in
healing ulcers. Phenolic compounds present in medicinal plants, have been reported to
possess powerful antioxidant activity1.
Herbal products are gaining a lot of demand these days because of their safety and efficacy.
They are more compatible and economical. Grape seed is a waste product in winery with 6090% procyanidine, which is potent antioxidant. Grape Seed Extract (GSE) has been shown to
possess various activities like antioxidant, antibacterial (Helicobacter Pylori, MethicillinResistant Staphylococcus Aureus), anticarcinogenic, antifungal, antihypolipidemic,
antihepatogenic 1, 2.
Since antiulcerogenic effect of GSE using various models simultaneously has not been done,
this study is undertaken to establish the same. Establishment of its antiulcer activity will
provide a newer and economically better modality of treatment for peptic ulcer, which is a
more common present-day ailment.
6.2 REVIEW OF LITERATURE:
Shi J et al of Korea, in 2003, have elaborately elicited the biochemistry and functions of
grape seeds. Grape seed possess lipid, protein, carbohydrates, and polyphenols depending on
the variety of grapes. Flavonols are major polyphenols of grapes. Flavanols have generated
remarkable interest based on positive reports of their antioxidant properties and ability to
serve as free radical scavengers. The beneficial effects of grape seed polyphenols is superior
to other well known antioxidants, such as vitamin C, vitamin E and beta carotene.
Antioxidant potency of proanthrocyanidins, a phenol, is 20 times greater than vitamin C and
50 times greater than vitamin E. The total extractable phenolics in grape are present at only
about 10% or less in pulp, 28-35% in the skin and 60-70% in the seeds1.
Maheswari U.M. of Coimbatore, in 2004, assessed the antihepatotoxic effect of grape seed oil
in rats. It was found out that oral administration of Grape Seed Oil in rats, significantly
decreased serum hepatic enzymes (malondialdehyde and hydroperoxide), which were
elevated after carbon tetrachloride (CCl4) treatment. Grape Seed Oil had no effect in control
rats. Hepatoprotective activity is attributed to the antioxidant mechanism which is brought
about by protecting against free-radicals2.
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Brown J. C. et al of America, in 2008, explored the alternate methods to treat Helicobacter
Pylori [H. Pylori] by determining its activity using agar dilution, confocal laser scanning
microscopy and cell proliferation assays following treatment with various grape extracts. The
study was done in-vitro and their activity against H. Pylori was established. The study
suggests that, higher phenolic levels do not necessarily determine overall anti-helicobacter
pylori activity. Their study showed Muscadine grape skin to possess the strongest activity,
followed by grape synergy (skin and seed) and seed3.
Sreemantula S et al of Vishakapattanam, in 2005, assessed the adaptogenic and nootropic
activities of grape seed. In their study, rats treated with grape seed, were subjected to forced
swim test. 24 hour excretion of urinary vanillylmandelic acid (VMA) and ascorbic acid was
assessed. Their levels were compared with that of normal rats. Comparison of their values
established the anti-stress (adaptogenic) action of GSE. The cognition, as determined by the
acquisition, retention and recovery, was found to be dose dependent. The conditioned
avoidance response analysis, established its nootropic activity. The antioxidant activity was
determined based on its ability to scavenge the hydroxyl radicals, as measured by studying
the competition between deoxyribose and the extract for hydroxyl radicals4.
Saito M et al of Japan, in 1998, assessed the gastroprotective activity of flavanols and
procyanidines present in grape seed in rats. They used grape seed having lower flavanol
content, higher flavanol content and procyanidins to assess their gastroprotective activity.
The dose used by them was 200mg/kg body weight. Ulcer was induced by 60% Ethanol
containing 150mM Hydrochloride. It was seen that the test compounds used strongly
inhibited the stomach mucosal injury. The mechanism of flavanol was by protection against
radical injury and of procyanidines by covering the gastric mucosal surface5.
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6.3 OBJECTIVES OF THE STUDY:
1. To demonstrate the Anti-Ulcer Activity of GSE using the following
Experimental Rat Ulcer Models:
A. Aspirin (200mg/kg) and Pylorus ligation model
with standard of Ranitidine (50mg/kg)
-Anti-secretory mechanism
B. Hydrochloric Acid [HCl] (0.3M) – Ethanol (60%) model
with standard of Sucralfate (100mg/kg)
-Cytoprotective mechanism
C. Water-immersion Stress model
with standard of Omeprazole (20mg/kg)
- Proton Pump Inhibition mechanism
2. To compare the anti-ulcer efficacy of GSE with that of respective standard drug.
3. To evaluate the efficacy of treatment using two doses (100mg/kg body weight,
200mg/kg body weight) of GSE.
4. To determine the probable anti-ulcer mechanism of GSE.
7. MATERIALS AND METHODS:
7.1 Source of Data:
Inbred Wistar Albino Rats of either sex, weighing 180 - 250 grams, from Institutional Animal
House, housed under standard conditions, caged separately.
7.2 Methodology of Study:
 Design and Sample size:
Three models, each model comprising of four groups, with each group having 6 rats,
72 rats in total.

Groups:
I.
II.
III.
IV.
Control(Gum Acacia)
Standard(STD) drug
GSE (100mg/kg body weight)
GSE (200mg/kg body weight)
o All drugs are administered orally.
o Rats will be sacrificed by decapitation.
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 Methodology:
o Model A:
Rats are fasted (water allowed), 200mg/kg Aspirin is given once for 3 consecutive
days. On the fourth day, test compounds are administered. After 30minutes, pylorus is
ligated under anaesthesia. Four hours later, rats will be sacrificed6,7.
o Model B:
Rats are fasted (water allowed) for 1day. Next day, test compounds are given. After
30mins, 1ml HCl and Ethanol are administered. 1hour later, rats are sacrificed7.
o Model C:
Rats are fasted for 1day (no water). Next day, test compounds are given. Rats will be
subjected to forced swimming in standard glass cylinder (height-45cms, diameter25cm) containing 35cms of water for 3 hours. Rats will be sacrificed.
 Analysis:
Stomach is excised; Contents-drained into centrifuge tube; Stomach - histologically
analysed.
 Statistical analysis employed:
ANOVA and Student t test

Outcome measures:
 Free and Total acidity
 Ulcer severity
 Ulcer index
 Does this study require any Investigations or Interventions to be
conducted on patients or other humans or animals? If so, describe
briefly
Yes.
Gross evaluation of ulcer number and ulcer size
Biochemical evaluation of free and total acidity
 Has ethical clearance been obtained from your institution:
Submitted to Institutional Animal Ethical Committee for approval
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8. List of references:
1. Shi J, Yu J, Pohorly J.E., Kakuda Y. Polyphenolics in Grape Seeds- Biochemistry and
Functionality. Journal of Medicinal Food. 2003; 6(4): 291-299.
2. Maheswari U.M., Rao P. G. M., Antihepatotoxic effect of grape seed oil in rat. Indian
Journal of Pharmacology. 2004; 37(3): 179-182
3. Brown J.C., Huang G, Haley-Zitlin V, Jiang X. Antibacterial effects of grape extracts
on Helicobacter pylori. Application of Environmental Microbiology. 2009; 75(3):84852.
4. Sreemantula S, Nammi S, Kolanukonda R, Koppula S, Boini K M. Adaptogenic and
nootropic activities of aqueous extract of Vitis vinifera (grape seed): An experimental
study in rat model. Biomed Central Complement of Alternate Medicine. 2005; 5:1.
5. Saito M, Hosoyama H, Ariga T, Kataoka S, Yamaji N. Antiulcer activity of Grape
Seed Extract and Procyanidins. Journal of Agricultural Food Chemistry. 1998; 46(4):
1460-1464.
6. Shay H, Komarov S A, Fels S E, Meraze D, Gruenstein M, Siplet H. A simple method
for the uniform production of gastric ulceration. Gastroenterology. 1945; 5 : 43–61.
7. Bickel M. Activity on gastrointestinal tract in Drug discovery and evaluation.
Pharmacological assays. Editors: Vogel G H, Vogel W H, Scholkens B A, Sandow J,
Muller G, Vogel W F. 2nd edition. Springer Publications. Germany. 2002: 867-869.
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Signature of the candidate:
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Remarks of the Guide:
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11.1 Name & Designation Dr. (Mrs.) VIJAYA RAJENDRAN. M.D.
of the Guide
Professor ,
Department of Pharmacology,
Vydehi Institute of Medical Sciences & Research Centre,
Bangalore 560066.
So far no study has been done on the antiulcer activity of grape
seed extract in India, which is a most welcoming study in
Bangalore.
11.2 Signature
11.3 Co-Guide
(if any)
11.4 Signature
11.5 Head of Department
Dr. B.N.SRINIVAS. M.D.
Professor & Head of Department,
Department of Pharmacology,
Vydehi Institute of Medical Sciences & Research Centre,
Bangalore 560066.
11.6 Signature
12.
12.1 Remarks of Chairman
& Principal
12.2 Signature
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