Wednesday 5 September 2012

ANTI-ULCER ACTIVITY OF ARISTOLOCHIA BRACTEOLATA


Pharmacologyonline 1: 1078-1082 (2011) ewsletter iyas et al.
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A TIULCER ACTIVITY OF AQUEOUS EXTRACTS OF ARISTOLOCHIA
BRACTEOLATA LEAVES
Mohamed iyas K ⃰ , Rupesh Kumar M, Tamizh Mani T, Fasalu Rahiman O.M, Surendra
Bodhanapu, Pasumarthi Phaneendra , Sathya Kumar B.
*Author for correspondence:
Mohamed Niyas. K
Dept. of Pharmacology,
Bharathi College of pharmacy,
Bharathinagara, Mandya,
Karnataka, India – 571422
Email: niaznasu@gmail.com
niyas_k97@yahoo.com
Summary
Peptic ulcer is a chronic and recurrent disease, and is the most predominant of the
gastrointestinal diseases. It is an excoriated area of the gastric or duodenal mucosa caused by
action of the gastric juice. This study focuses on the anti ulcer activity of aqueous extract of
the plant Aristolochia bracteolata, a perennial herb, the leaves of which are used by the native
tribals and villagers of the Chittoor District of Andhra Pradesh in India for the rapid healing
of cuts and wounds. The aqueous extract of the shade dried leaves of Aristolochia bracteolata
was studied for its antiulcer activity in rats, using ethanol induced and pylorus ligation
induced models, at two different dose levels of 400 and 800 mg/kg/body wt/day. The activity
was compared with standard drug Ranitidine. Pretreatment with the extract resulted in a
significant decrease of the ulcerated area. The volume and acidity of the gastric juice
decreased in the pretreated rats. Among the two dose assessed, 800 mg/kg was found to have
the significant activity than the lower dose.
Key words: Aristolochia bracteolata, antiulcer activity, aqueous, ranitidine.
Abbreviations: Aqueous extract of Aristolochia bracteolata (AEAB)
Introduction
Peptic ulcers are a deep gastrointestinal erosion disorder that involves the entire mucosal
thickness, penetrating the muscular mucosa [1]. For decades it was believed that
gastrointestinal ulcerations were caused by the excessive secretion of gastric acid, but many
patients presenting such ulcerations had normal acid secretion rates. Then, researchers
reported that peptic ulcers were been caused by an imbalance between the aggressive factors
and a number of known defense mechanisms [2]. It is a chronic and recurrent disease, and is
the most predominant of the gastrointestinal diseases [3].
Pharmacologyonline 1: 1078-1082 (2011) ewsletter iyas et al.
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Aristolochia bracteolata is a shrub distributed throughout India. It belongs to the family
Aristolochiaceae. In the indigenous system of medicine, the plant was used for the treatment
of skin diseases, inflammation and purgative [4]. Root extract was reported to have anti
bacterial activity [5]. It has insecticidal properties. Its leaves are bitter and antihelmintic, and
are medicinally important. Almost every part of the plant has medicinal usage. Aristolochia
bracteolata is proved to have antioxidant property [6]. The antiulcer activity of Aristolochia
bracteolata has not yet been studied. Hence the aim of the present investigation was to
evaluate the antiulcer activity of aqueous extract of Aristolochia bracteolata leaves.
Materials and methods
Plant material: The fresh leaves of Aristolochia bracteolata was collected from Tirupati,
Chittoor district, Andhra Pradesh. The plant was identified, confirmed and authenticated by
comparing with voucher specimen available at Calicut university herbarium, Department of
botany, university of Calicut, Emerald by Botanist Dr. Pradeep AK and voucher specimen
was deposited in institutional herbarium.
Preparation of extract: The fresh leaves of Aristolochia bracteolata was dried under shade.
The dried leaves were powdered using a grinder. The coarse powder was used for extraction.
Powdered leaves of Aristolochia bracteolata was extracted by maceration technique for 7
days.
Animals: Healthy adult albino rats of Wistar strain weighing 150-200g of either sex were
used for this study. The animals were obtained from animal house, Bharathi College of
Pharmacy, Bharathinagara, Karnataka, India. The animals were maintained under controlled
conditions of temperature (23 ± 2°C), humidity (50 ± 5%) and 12-h light-dark cycles. All the
animals were acclimatized for seven days before the study. The animals were randomized into
experimental and control groups and housed individually in sanitized polypropylene cages
containing sterile paddy husk as bedding. They had free access to standard pellets as basal
diet and water ad libitum. Animals were habituated to laboratory conditions for 48 h prior to
experimental protocol to minimize if any of non-specific stress.
Experimental Design
a) Ethanol induced ulcers:
Four groups of albino Wistar rats (n=6) were selected. In this model, Group 1 served as
normal control received 0.5 ml of vehicle, p. o., and group 2 received Ranitidine (80 mg/kg,
p.o), whereas groups 3 and 4 animals received aqueous extract of Aristolochia bracteolata
(400 and 800 mg/kg, p.o. respectively). Animals were fasted overnight prior to start of the
experiment, and water ad libitum 30 min after treatment, all rats received 1ml of absolute
ethanol to induce gastric ulcer. After 1 h the animals were sacrificed by cervical dislocation,
the stomachs were removed and opened along the greater curvature. Stomachs were gently
rinsed with water to remove gastric contents and the mean ulcer index was calculated [8].
b) Pylorus ligation induced ulcers:
Four groups of albino Wistar rats (n=6) were selected. In this model, Group 1 served as
normal control received 0.5 ml of vehicle, p. o., and group 2 received Ranitidine (80 mg/kg,
p.o), whereas groups 3 and 4 animals received aqueous extract of Aristolochia bracteolata
(400 and 800 mg/kg, p.o. respectively). Animals were fasted overnight prior to start of the
experiment, and water ad libitum Pyloric ligation was applied by ligating the pyloric end of
the stomach of rats under Phenobarbital anaesthesia ( 35 mg/kg) after 30 min of aqueous
Pharmacologyonline 1: 1078-1082 (2011) ewsletter iyas et al.
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extract of Aristolochia bracteolata or ranitidine treatments. Animals were allowed to recover
and stabilize in individual cage and were deprived of water during postoperative method.
After 6 h of surgery, rats were sacrificed with excess ether and gastric juice was collected for
performing gastric secretion study and ulcer scoring was done in stomach [7].
Statistical analysis: The data of results obtained were subjected to statistical analysis and
expressed as mean ± SEM. The data were statistically analyzed by one-way analysis of
variance (ANOVA) and p<0.01 was considered to be significant and p<0.001 was considered
to be more significant.
Results and discussion
Ethanol induced ulcer model
The effect of Aristolochia bracteolata on ethanol induced gastric ulcers is given in table no.1
and fig. no.1 to fig. no. 4. Ethanol (80%, 1 ml) induced ulcers in normal control animals was
evidenced by the ulcer index (UI) 13.117 ± 0.641, high acid volume 8.43 ± 0.084 ml, low pH
2.1 ± 0.118, high total acidity 111.5 ± 1.176 mEq/l, low glutathione 0.418 ± 0.009 μg/g and
high total protein 0.372 ± 0.012 g/dl. When aqueous extracts of Aristolochia bracteolata was
given along with ethanol (80%, 1 ml) at two dose levels, 400 mg/kg and 800 mg/kg b.w.
caused a significant reversal of all the above parameters when compared to control rats
indicating its potent antiulcer activity. Among the two doses Aristolochia bracteolata at 800
mg/kg b.w. was found to be more effective than the lower dose.
Table o.1 Effect of A.bracteolata on ethanol induced gastric ulcers
Parameters
No. Treatment
Dose
mg/kg Ulcer
Index
Acid
Volume
(ml)
pH
Total
acidity
(mEq/l)
Glutathione
(μg/gm)
Total
Protein
(gm/dL)
1
control
80%
13.117±
0.641
8.43±
0.084
2.1±
0.118
111.5±
1.176
0.418±
0.009
0.372±
0.012
2 Standard 80
4.167±
0.154***
2.68±
0.079***
6.067±
0.14***
58.67±
0.72***
0.892±
0.012***
0.15±
0.01***
3
AEAB
400
11.63±
0.102*
8.00±
0.146*
2.9±
0.139*
107.16±
0.792*
0.457±0.02
0.368±
0.009
4 AEAB 800
11.317±
0.209**
7.93±
0.084**
3.08±
0.164**
106.33±
1.02**
0.523±0.023*
0.325±
0.012*
Values are expressed as Mean ± SEM from 6 rats.P<0.01** and P<0.001*** as compared to
control group.
Pharmacologyonline 1: 1078-1082 (2011) ewsletter iyas et al.
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Pylorus ligation induced ulcers:
The effect of Aristolochia bracteolata on pylorus ligation induced gastric ulcers is given in
Table No.2. Pylorus ligation induced ulcers in control animals are evidenced by the ulcer
index 11.717 ± 0.179, high acid volume 8.683 ± 0.654 ml, low pH 2.53 ± 0.076, high total
acidity 41.67 ± 0.615 mEq/l, low glutathione 0.383 ± 0.048 μg/g and high total protein
0.367± 0.022 g/dl. When aqueous extracts of Aristolochia bracteolata was given at two dose
levels, 400 mg/kg and 800 mg/kg b.w. caused a significant reversal of all the above
parameters when compared to normal control rats indicating its potent antiulcer activity.
Table o.2 Effect of A.bracteolata on pylorus ligation induced gastric ulcers
Parameters
No. Treatment
Dose
mg/kg Ulcer
Index
Acid
Volume
(ml)
pH
Total
acidity
(mEq/l)
Glutathione
(μg/gm)
Total
Protein
(gm/dL)
1
control
(Ethanol)
80%
11.717±
0.179
8.683 ±
0.654
2.53 ±
0.076
41.67±
0.615
0.383±
0.048
0.367±
0.022
2 standard 80
3.283 ±
0.13***
2.32 ±
0.075***
6.417±
0.08**
*
11±
0 .577***
1.017±
0.079***
0.13±
0.02***
3
AEAB
400
11.05±
0.163**
8.467±
0.062
3.0±
0.085*
40.16±
0.946
0.533±0.033
0.317±
0.031
4 AEAB 800
9.4±
0.097***
8.267±
0.076**
3.05±0.
134**
37.5±
0.563**
0.7±0.026*
0.217±
0.031*
Values are expressed as Mean ± SEM from 6 rats.P<0.05*, P<0.01** and P<0.001*** as
compared to normal control group.
Effect of A.bracteolata on ethanol induced gastric ulcers
Fig.1 Control (Ethanol) Fig.2 Standard (Ranitidine)
Pharmacologyonline 1: 1078-1082 (2011) ewsletter iyas et al.
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Fig.3 AEAB (400 mg/kg) Fig.4 AEAB (800 mg/kg)
Conclusions
From the data of results obtained it is evaluated that aqueous extract of the plant Aristolochia
bracteolata possesses a significant antiulcer activity compare to the standard drug. The study
also helped us to identify the therapeutic values of the common plants present around us.
Acknowledgements
The author is thankful to, Bharathi College of pharmacy, Bharathinagara, Karnataka, India for
providing necessary facilities throughout this work.
References
1. Tarnawski AS. Cellular and molecular mechanisms of gastrointestinal ulcer healing.
Digest Dis Sci 2005; 50: 24-33.
2. Wallace JL, Granger DN. The cellular and molecular basis of gastric mucosal defense.
FASEB J 1996; 10: 731-740.
3. Guyton and Hall. Textbook of Medical Physiology, 10, Philadelphia, 2000; 397-398.
4. Wealth of India, Raw Materials Vol. IA, CSIR, New Delhi, 1982; 88.
5. Negi, P.S.,Anantharamakrishnan,C, Jayaprakasha,G.K. J Med Food 2003; 6: 401.
6. Shirwaikar A, Somashekar AP, Antiinflammatory activity and free radical scavenging
studies of Aristolochia bracteolata Lam. Indian J. Pharm Sci 2003: 67-69.
7. Vogel HG, Drug Discovery and Evaluation, 2, Spinger-Verlag Berlin Heidelberg,
New York; 2002: 867-71.
8. Hussain R, Turaifi A, Ahmed AM, Effect of Clotrimazole on Chemically and Stress
Induced Peptic Ulcer, Scientific Journal of King Faisal University 2007; 8: 1-15.

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