1 Received by Belinda Received on 2012-7-16
2 ID No. B541 Revised on 2012-7-27
3 Pages 13
5 Evaluation of Antiulcer Activity of Bark Extract of Albizzia lebbeck Linn.
6 Neelam Balekar*, Dinesh Kumar Jain, Hitesh Jawanjal.
7 College of Pharmacy IPS Academy, Rajendra Nagar, A.B. Road, Indore – 452012
8 (M.P.), INDIA
13 *Corresponding author: Dr. Neelam Balekar,
14 Tel.: +919893405071; fax: +07314041627.
15 E-mail address: firstname.lastname@example.org (Neelam Balekar).
16 Postal Address: Department of Pharmacology, College of Pharmacy, IPS Academy,
17 Hukhmakedhi, Rajendra Nagar, A.B. Road, Indore – 452012 (M.P.),
6 Objective: To determine the antiulcer properties of the ethanolic extract from the stem
7 bark of Albizzia lebbeck Linn. in experimental rats.
8 Methods: Antiulcer potential of ethanolic stem bark extract of A. lebbeck were
9 determined using ethanol induced, aspirin induced, cold stress restraint and pylorus
10 ligated ulcer in rats.
11 Results: The ethanol extract (100, 250, 500,and 1000 mg/kg, p.o.) significantly
12 suppressed the peptic ulcer induced by ethanol. Ethanolic extract (500 mg/kg) showed
13 promising activity. Hence, this dose was selected for the further evaluation of antiulcer
14 studies. The ethanolic extract (500 mg/kg) showed significant (P< 0.05) reduction in
15 gastric volume, free acidity, total acidity, ulcer index, protein and pepsin content and
16 increase in mucus content as compared to control.
17 Conclusions: The present study indicates that A. lebbeck bark extract have potential
18 antiulcer activity in all models which might be due to its antisecretory activity and
19 increased resistance to necrotizing agents, providing a direct, protective effect on the
20 gastric mucosa.
21 Keywords: Albizzia lebbeck, Gastric ulcer, cytoprotection, antisecretory.
1 1. Introduction
2 Gastric ulcer, most common disorder of gastrointestinal tract has multifunctional causes
3 in its pathophysiology. The disorder affects large population in all countries. It is now
4 assumed that these drugs ultimately balance the aggressive factors (acid, pepsin, H.
5 pylori, bile salts) and defensive factors (mucin secretion, cellular mucus, bicarbonate
6 secretion and mucosal blood flow). Peptic ulcer is a lesion of gastric or duodenal
7 mucosa occurring at site where the mucosal epithelium is exposed to aggressive
8 factors. Herbal medicine is fast emerging as an alternative treatment to available
9 synthetic drugs for treatment of ulcer possibly due to lower costs, availability, fewer
10 adverse effects and perceived effectiveness. Albizzia lebbeck Linn. (family-
11 Mimosaceae) is a tropical evergreen tree widely distributed in India, South Africa and
12 Australia. Its stem bark has been used for a long time in Southeast Asia as a traditional
13 medicine for toothache, diseases of gum, skin infection, ulcer, piles, dysentery and
14 diarrhea[3,4]. A. lebbeck contains tannin, flavonoids, steroids, and triterpenoids. The
15 pharmacological activities previously reported are antibacterial, antiinflammatory,
16 antimicrobial, and antioxidant.
17 The aim of the present study was to investigate the antiulcer activitiy of the bark of
18 A. lebbeck commonly employed by traditional healers. Parameters like ethanol induced,
19 aspirin induced, cold restraint stress and pylorus ligated ulcer model in rat were utilized
20 to clarify its traditional use in a scientific investigation.
22 2. Materials and Methods
23 2.1 Plant material and chemicals
1 The fresh stem bark of A. lebbeck Linn. (Mimosaceae) was collected in October, 2009
2 from A.B. Road, Indore (M.P.). The plant was authenticated and voucher specimen
3 (ALHIT1) was deposited at the herbarium of the Botanical survey of India, Pune.
4 Aspirin was obtained as gift sample from Cyno Pharma, Indore, India and omeprazole
5 and ranitidine were obtained from Alpa Laboratories Indore, India. All the other
6 chemicals and reagent used were of analytical grade, obtained from Kasliwal Brothers,
7 Indore, India.
9 2.2 Extraction
10 The stem bark (500 g) of A. lebbeck was shade dried, coarsely powdered and defatted
11 by maceration with petroleum ether for 48 h. The defatted marc was subjected to soxhlet
12 extraction with 95% ethanol (2.5 L) at (60-80 ºC). The solvent was filtered through
13 Whatman No. 1 filter paper and evaporated to dryness under vacuum at 40 ºC using
14 rotary evaporator. The ethanolic extract was stored in tightly closed container and kept
15 in refrigerator.
17 2.3 Preliminary phytochemical screening
18 The extract was qualitatively analyzed for the presence or absence of phytoconstituents
19 like glycosides, flavonoids, saponins, alkaloids, carbohydrates, sterols, proteins using
20 standard methods. The quantitative determination of total flavonoid content was done
21 by the method of Chang et al., 2002.
1 2.4 Animals
2 The Wistar rats of either sex weighing 120-200 g were used. Animals were housed
3 under standard condition of temperature (25 ± 2 ºC), 12 h/12 h light/dark cycle and fed
4 with standard pellet diet and water ad libitum. The animals were allowed to acclimatize
5 for one week before the experiments. All experimental protocols were approved by the
6 IAEC under the supervision of CPCSEA.
8 2.5 Ethanol induced ulcer
9 Wistar rats were divided into six groups, each group containing five animals. Group I
10 was control (0.5% Sodium carboxymethyl cellulose (SCMC)), group II served as
11 standard (Omeprazole 20 mg/kg p.o.), group III-VI served as ethanolic extract ALEE
12 (100, 250, 500, 1000 mg/kg) were administered to rats 1 h before ethanol (5 ml/kg)
13 treatment. After 1 h, the animals were sacrificed by cervical dislocation and the gastric
14 lesions were counted.
16 2.6 Aspirin induced ulcer
17 Wisar rats were divided into three groups. Group I was control (0.5% SCMC), group II
18 served as standard (ranitidine 50 mg/kg), group III was ALEE (500 mg/kg). Each group
19 was containing five animals. All treatments were administered 30 min before aspirin.
20 Aspirin (1000 mg/kg) was administered orally to rats. The animals were sacrificed by
21 cervical dislocation 6 h later and the stomach was removed to calculate the ulcerative
24 2.7 Cold stress restraint ulcer
1 Wistar rats were divided into four groups. Group I was positive control (restraint with
2 cold stress), Group II served as negative control (restraint without cold stress),
3 Group III served as standard (omeprazole 20 mg/kg). Group IV was ALEE (500
4 mg/kg). Each group was containing five animals. All treatments were administered 1 h
5 before stress in restraint cages that were kept at 2 ºC in chamber for 4 h treatment. After
6 4 h, the animals were sacrificed using cervical dislocation and the gastric lesions were
7 counted as earlier and mucus content was estimated.
9 2.8 Pylorus ligated ulcer
10 Wisar rats were divided into three groups. Group I was control (0.5 % SCMC), group
11 II served as standard (omeprazole 20 mg/kg), group III was ALEE (500 mg/kg). Each
12 group was containing five animals. All treatments were administered to animals 45
13 mins before pylorus ligation. Pylorus ligation was done by ligating the pyloric end of
14 stomach of rats of respective groups under ether anaesthesia. Ligation was done
15 without causing any damage to the blood supply of the stomach. Animals were allowed
16 to recover and stabilize in individual cages and were deprived of water during
17 postoperative period. After 6 h of surgery, rats were sacrificed by overdose of ether and
18 ulcer scoring was done. Gastric juice was collected and mucus content, protein, pepsin
19 content, free and total acidity, pH were determined.
21 2.9 Statistical analysis
22 Data are expressed as a mean ± S.D. Statistical evaluation was carried out using one-
23 way (ANOVA followed by Tukey’s test) using “Graphpad Instat” version 3.00 for
1 Windows 95, Graphpad software, San Diego California USA. The values of P < 0.05
2 were considered to be statistically significant.
5 3.1 Phytochemical analysis
6 The percentage yield of the ethanolic extract was found to be 12.36%. The ethanolic
7 extract showed the presence of flavonoid, tannin, saponin, protein. The total flavonoids
8 content in ethanolic was found to be 12.15 mg quercetin equivalents/g.
10 3.2 Ethanol induced ulcer
11 Oral administration of the ethanolic extract of the stem bark of Albizzia lebbeck (100,
12 250, 500, and 1000 mg/kg, p.o.) dose dependently reduced the ulcer index induced by
13 ethanol in rats as compared to control (P < 0.01). The mean ulcer index was found to be
14 control, (22.12 ± 0.74), omeprazole 20 mg/kg (2.1 ± 0.74) and ALEE 500 and 1000
15 mg/kg, (3.3 ± 0.84, 3.1 ± 0.89), respectively. At the dose of 100, 250, 500 and 1000
16 mg/kg, ulcer index was inhibited by 37.16, 69.71, 85.08 and 85.99% respectively. The
17 reference drug, omeprazole (20 mg/kg), also produced a significant protective effect
18 towards the ethanol induced ulcer by 90.51% inhibition (Fig 1). The reduction in ulcer
19 index produced by ALEE 500 and 1000 mg/kg was found to be comparable to
22 3.3 Aspirin induced ulcer
23 The ethanolic extract of A. lebbeck stem bark (500 mg/kg, p.o.) and ranitidine
24 (50 mg/kg, p.o) significantly exert protective effect on aspirin induced ulcer in rats. The
1 ulcer protective effect exhibited by A. lebbeck and standard ranitidine was found to be
2 82.43 % and 89.86% respectively (Fig 2). The reduction in ulcer index produced by
3 ALEE 500 mg/kg (2.6 ± 0.65) was found to be comparable to ranitidine (1.5 ± 0.5).
5 3.4 Cold restraint stress ulcer
6 The ethanolic extract of A. lebbeck stem bark (500 mg/kg, p.o.) and omeprazole
7 (20 mg/kg, p.o) significantly exert protective effect on cold restraint stress ulcer in rats.
8 The mean ulcer produced by ALEE 500 mg/kg was significantly reduced as compared
9 to positive control (P<0.05). The ulcer protective effect exhibited by A. lebbeck and
10 standard omeprazole was found to be 78.46% and 85.93 % respectively (Table 1).
12 3.5 Pylorus ligated ulcer
13 In pyloric ligation induced ulcer model, oral administration of ethanolic extract
14 (500 mg/kg) showed significant reduction in ulcer index, gastric volume, free acidity,
15 total acidity as compared to the control group (P<0.05). It showed protection index of
16 77.89 % at the dose of 500 mg/kg in comparison to control whereas omeprazole as
17 reference standard drug showed reduction of ulcer by 85.26 % (Table 2).
19 3. Discussion
20 The antiulcer activity of the bark of A. lebbeck was evaluated by employing ethanol,
21 aspirin, cold restraint stress and pylorus ligation ulcer models. These models represent
22 some of the most common causes of gastric ulcer in humans. Ulcers caused by
23 ethanol are due to superficial damage to mucosal cells and damage by NSAIDs are
24 due to a decrease in PG synthesis, and ulcers due to stress are due to both physiological
1 and psychological factors and those by pyloric ligation are due to increased
2 accumulation of gastric acid and pepsin leading to auto digestion of the gastric
4 Antiinflammatory drugs like aspirin administered in toxic doses (1000 mg/kg), produce
5 visible gastric ulcers in animals. Aspirin is a potent inhibitor of prostaglandin
6 biosynthesis. Prostaglandins are known to play an important role in maintaining
7 mucosal integrity. An increase in certain endogenous prostaglandins can enhance
8 gastric mucosal resistance to ulcerogenic agents. The mechanisms involved in
9 prostaglandin action are multiple, including stimulation of mucus and bicarbonate
10 output, gastric mucosal blood flow that may be due to the presence of flavonoid
11 in A. lebbeck extract.
12 Ethanol induced gastric ulcer was employed to study the cytoprotective effect of the
13 extracts. Ethanol challenge induces gastric injury due to production of oxygen free
14 radicals leading to increased lipid peroxidation, which causes damage to cell and cell
15 membrane presenting as red streaks of sores. Ethanol induced gastric lesion
16 formation may be due to stasis in gastric blood flow which contributes to the
17 development of the haemorrhage and necrotic aspects of tissue injury. A. lebbeck
18 extracts has significantly decrease the ulcer index, may be due to the presence of
19 flavonoid which may showed antisecretory activity. Omeprazole were employed in this
20 study for the latter’s cytoprotective but not anti-secretory effect and its effectiveness
21 against experimentally induced ethanol ulcers.
22 On the other hand, pyloric ligation induced ulcers develop as a result of accumulation
23 of the gastric acid and distention of the stomach which in turn weakens the mucosal
24 defenses. It has been proposed that in pyloric ligation, the digestive effect of
1 accumulated gastric juice and interference of gastric blood circulation are responsible
2 for induction of ulceration.
3 In the cold-restraint stress model, gastric ulcer formation was mainly due to gastric
4 hypermotility, which could lead to mucosal over friction. Hence, the gastric mucus
5 layer is extremely important and the mucus is generally believed to contribute to a
6 cytoprotective action. In the stomach, prostaglandin synthase is the major pathway,
7 since leukotriene synthesis is approximately half of prostaglandin synthesis. Stress
8 induces an increase in leukotriene synthesis in the stomach which, compared to PGE2,
9 indicates that arachidonate oxidation changed toward leukotriene production. The
10 ethanolic extract of A. lebbeck (500 mg/kg, p.o.) showed significant reduction in cold
11 restraint stress in rats
12 The possible mechanism of gastric mucosal protection by ALEE may be partially due
13 to the reinforcement of resistance of the mucosal barrier by a protective coating. The
14 ethanolic extract of A. lebbeck in concentration dependent manner exhibited marked
15 gastroprotection in ethanol and aspirin induced ulcer model. The present investigation
16 thus, establishes the ulcer protective and healing effects of ALEE in different gastric
17 ulcer models and activity seemed to be due to its effect both on offensive and defensive
18 mucosal factors.
20 Conflict of interest statement
21 We declare that we have no conflict of interest.
1 The authors would like to acknowledge College of Pharmacy, IPS Academy, Indore,
2 India for providing the necessary facilities to carry out the study.
4 Research grant information;
5 SOURCE: This work was supported by IPSA Research Devlopment Scheme
6 GRANT NO- IPSA/RDS/2009-10/02
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