Vol. 7(34), pp. 2537-2541, 10 September, 2013

DOI: 10.5897/JMPR2013.4441

ISSN 1996-0875 ©2013 Academic Journals

http://www.academicjournals.org/JMPR

Journal of Medicinal Plants Research

Full Length Research Paper

Anti-ulcer activities of oil extract of Balanites aegyptiaca seed in guinea pigs

C. A. Eze1, D. C. Anyogu2*, I. C. Nwogu3 and S. S. Adamu4

1Department of Veterinary Surgery, University of Nigeria, Nsukka, Nigeria.

2Department of Veterinary Pathology, University of Nigeria, Nsukka, Nigeria. 3Department of Veterinary Anatomy, University of Nigeria, Nsukka, Nigeria.

4Department of Veterinary Surgery and Reproduction, University of Maiduguri, Nigeria.

Accepted 27 August, 2013

Over the years, management of advanced peptic ulcers used to be a nightmare and in most cases surgery is resorted to. This study was carried out to investigate the anti-ulcer activities of the oil extract of Balanites aegyptiaca seed in guinea pigs. Thirty guinea pigs weighing between 200 and 440.8 g were randomly assigned into six groups of five each. Gastroduodenal ulceration was achieved by pyloric ligation and histamine injection. The anti-ulcer effect of the oil was compared with omeprazole (10 mg/kg body weight) given orally. The oil extract of B. aegyptiaca seed did not significantly reduce the mean severity of ulcer incidence in both pyloric ligation and in histamine injection ulcer models. But it significantly (P≤0.05) decreased the volume of gastric secretion in histamine injection model. It equally decreased the percentage of ulcer incidence in the guinea pigs and raised the percentage protection in both models of experiment by 40.9 and 50.6%, respectively. However, omeprazole at the dose rate of 10 mg/kg body weight gave better protection of 80.3 and 84.4% against ulcer in both models, respectively. The results showed that B. aegyptiaca oil possessed moderate gastric anti-secretory and cytoprotective properties which can be employed in the prevention, treatment and relief of gastric ulcer symptoms. Key words: Anti-ulcer, Balanites aegyptiaca, guinea pig, pyloric ligation, histamine.

INTRODUCTION Peptic ulcer is caused by an uncontrollable increase in gastric acid secretion from the gastric glands, which are located in the body of the stomach. It occurs as a breach in the mucosa of the alimentary tract that extends through the muscularis mucosa into the submucosa or deeper (Crawford, 1997). Many chemotherapeutic agents have been employed over the years for the management of peptic ulcer, although surgery may be required in advanced cases. Following the discovery of Helicobacter pylori as the aetiologic agent of peptic ulcer disease, a triple therapy regimen was recommended for gastric

hyper-secretors to include a mixture of antacids, anti-secretory agents and antibiotics (Tan and Nyasse, 2000). The control of H. pylori as well as H

+/K

+- adenosine

triphosphatase (ATPase), acid secretion and subsequent reversal of mucosal damage and inflammation have been utilized in the current therapy for this disease (Woolf et al., 2002). This triple therapy regimen is difficult to obtain in Africa, and almost unavailable especially to the rural poor, triggering off the search for cheaper and easily available anti-ulcer agents amongst natural products-herbs, fish and plant oils (Goel and Sairam, 2002).

*Corresponding author. E-mail: davinson.anyogu@unn.edu.ng.

2538 J. Med. Plants Res.

Balanites aegyptiaca (Desert date) oil, cooking oil in Nigeria, has been found to speed-up surgical wound healing, probably due to its antimicrobial and soothing effects (O’Leary and Capote, 1996; Harvey et al., 1990; Hillman and Bishop, 1996; Aiello and Mays, 1998; Eze et al., 2006). Some investigators have also reported that B. aegyptiaca oil exhibited anticancer activity against lung, liver and human brain carcinoma cell lines in addition to its antimutagenic activity against Facsiola gigantica-induced mutagenicity (Al Ashaal et al., 2010). It equally has anthelmintic activity against hepatic worms (Schistosoma mansoni and Fasciola gigantic), antiviral activity against Herpes simplex virus as well as antibacterial activity against selected strains of Gram-positive and Gram-negative bacteria and candida (Al Ashaal et al., 2010). This study therefore, investigated the anti-ulcer properties of B. aegyptiaca oil in guinea pigs. MATERIALS AND METHODS Extraction of B. aegyptiaca kernel oil Fruits of B. aegyptiaca plant were obtained from Kwaya Town in Kwayah Kusar Local Government Area of Bornu State, Nigeria. Extraction of oil from seeds of B. aegyptiaca fruits was done locally using the manual method. Five kilograms of the kernel seed was soaked for 12 h in cold water. The kernels were then de-hulled to yield the seeds, which were roasted in an open pan and ground to paste.

Water (70 to 80°C) was poured on the seeds and the seeds were squeezed for hours to yield 100 ml of oil. The oil was then heated to remove any remaining traces of water and to maintain the quality and prevent water catalysis of oil rancidity. Animals Thirty guinea pigs, mean weight 300.0±7 g were randomly assigned into 6 groups of 5 animals each. Feed (standard commercial feed) and water were provided ad libitum. The guinea pigs were, however, fasted for 24 h with free access to drinking water prior to ulcer induction. Experiment I: Pyloric ligation method Guinea pigs in groups I, II and III were used in this method. Group I received no treatment prior to pyloric ligation. Group II received B. aegyptiaca oil, 1 ml per os, 1 h prior to pyloric ligation. Group III received omeprazole (10 mg/kg body weight peritoneally (p.o.)) 1 h prior to pyloric ligation. Pyloric ligation was achieved by laparotomy incision below the xyphoid process under chloroform anaesthesia as described by Shay et al. (1945). The incisions were sutured and the guinea pigs were kept for 4 h after which they were humanely sacrificed and the ulcer parameters scored. Experiment II: Histamine injection method

Group IV received no treatment prior to histamine injection while

groups V and VI received B. aegyptiaca oil (1 ml) and omeprazole (10 mg/kg body weight), p.o, 1 h prior to histamine injection, respectively. Histamine was administered at a dose of 0.25 mg/kg, body weight, intraperitoneally, at intervals of 30 min for 7 consecutive times to induce 100% gastro-duodenal ulceration in the guinea pigs (Watt and Eagleton, 1964).

The gastric lesions and the stomach contents in both experiments were harvested and the ulcer index scored as described by Hano et al. (1976), with a little modification. Measurement of the volume of gastric secretion The stomach contents of each guinea pig was harvested into a container and immediately filtered into a calibrated test tube using the Whatman filter paper. The volume of gastric secretion was then read off. Determination of ulcer parameters The following arbitrary scoring system was used to grade the severity and intensity of the lesions. Shedding of epithelium = 10; Petechial and Frank haemorrhages = 20; one or two ulcers = 30; more than two ulcers = 40; perforated ulcer = 50.

The presence of any of these lesions was considered a positive ulcerogenic response which was shown as percentage of guinea pigs showing gastric lesions. The severity of ulcers was expressed in terms of ulcer index, which is the mean score of gastric lesions of all the guinea pigs in a group.

Ulcer index (U.I) = US + UP × 10-1

where Us is mean severity of ulcer score and Up is the percentage of animals with ulcer incidences.

Percentage protection = (UC – UT / UC) × 100/1 where UC is the ulcer index of negative control and UT is the ulcer index of positive control or any treatment group. Histopathology Tissue sections of the stomach and duodenum were fixed in Bouin’s fluid, embedded in paraffin, sectioned at 5-µm thickness, routinely processed and stained with hematoxylene and eosine.

Statistical analysis Results were presented as mean and standard error (mean ± standard error (SE)). The significance between the control and each of the oil- and omeprazole-treated groups was determined by analysis of variance (ANOVA). The variant means were separated using the least significant difference method (LSD). The level of significance was set at P≤0.05. RESULTS Pyloric ligation as well as histamine injection induced 100% gastric ulcer in guinea pigs in the control groups.

Eze et al. 2539

Table 1. Parameters used to evaluate the extent of ulcer in pyloric ligation method.

Ulcer parameter Control (Group I) B. aegyptiaca oil (Group II) Omeprazole (Group III)

Mean severity of ulcer score 62.00±21.54a 24.00±14.69

ab 8.00±8.00

b

Percentage of ulcer incidence 100 (5/5) 60 (3/5) 20 (1/5)

Ulcer index 14.20 8.40 2.80

% protection 00 40.85 80.28

Volume of gastric fluid (ml) 6.54±0.86a 3.96±1.44

ac 0.82±0.37

b

Superscript = Significant difference at P≤0.05.

Table 2. Parameters used to evaluate the extent of ulcer in histamine injection method.

Ulcer parameter Control (Group IV) B. aegyptiaca oil (Group V) Omeprazole (Group VI)

Mean severity of ulcer score 58.00±31.37a 18.00±12.00

ab 4.00±4.00

abc

Percentage of ulcer incidence 100 (5/5) 60 (3/5) 20 (1/5)

Ulcer index 15.80 7.80 2.40

% protection 00 50.60 84.80

Volume of gastric fluid(ml) 4.50±0.58a 1.10±0.37

b 1.00±0.35

bc

Superscript = Significant difference at P≤0.05

However, pyloric ligation appeared to produce a severer ulceration resulting in a perforated ulcer in one of the guinea pigs in the control group. B. aegyptiaca-treated groups showed moderate reduction in the ulcer mean severity scores, although these were not statistically significant in both experimental procedures used to achieve gastric ulceration. The oil extract also reduced the percentage of ulcer incidence in the guinea pigs, the calculated ulcer index and the mean volume of gastric secretions, especially in histamine injection method (Table 2). Histologically, B. aegyptiaca oil allowed in the guinea pigs only a little digestion of the stomach wall by gastric acids leading to a relatively lesser destruction of the gastric mucosal epithelium and lamina propria (Figure 2) compared to the untreated groups.

Omeprazole (10 mg/kg body weight), however, gave a greater reduction in the results of the aforementioned parameters in both experimental protocols used to assess the ulcer lesions (Tables 1 and 2). As a result, only a very mild lesion was observed in the gastric mucosae of omeprazole-treated rats at histology (Figure 3). DISCUSSION Pyloric ligations, intra-peritoneal administration of hista-mine as well as cold-restraint methods produce gastric lesions in the glandular part of the stomach (Levine, 1965; Ogle et al., 1985). In this study, ulcer genesis in pyloric ligation was due to the accumulated gastric juice which caused stomach distention leading to an increase

Figure 1. Mucosal fold section of stomach of guinea pig from the control group. Note severe necrosis and loss of lining epithelium, glands and lamina propria (arrow) H&E 100×.

in histamine secretion (Tan et al., 2000). The acidity led to auto digestion of the gastric mucosae by the accumulated hydrochloric acid and pepsin (Figure 1). B. aegyptiaca oil decreased the volume of gastric secretion, though not significantly, in pyloric ligation induced ulceration, but very significantly in histamine induced gastric ulceration when compared with the control. This difference in the volume of gastric secretion can be attributed to differences in methods of achieving ulcers; the occlusion of pylorus does not allow gastric emptying

2540 J. Med. Plants Res.

Figure 2. Stomach section of guinea pig treated with Balanites aegyptiaca oil before inducing ulcer showing moderate necrosis of lining epithelium and lamina propria (arrow) H&E 100×.

Figure 3. Stomach section of guinea pig treated with omeprazole before inducing ulcer. Note mild excoriation (arrows) on lining epithelium. H&E 100×.

unlike in histamine injection in which gastric secretions flow down the duodenum. Agents that decrease gastric secretion are helpful in reducing ulcers due to stress (Olatunji-Bello, 2006).

The aforementioned reduction in ulcer parameters is in consort with the findings of Hasan (2004) and Grant (1990), which reveal that polyunsaturated fatty acids possess anti-ulcer and anti-oxidant effects. B. aegyptiaca oil is rich in linoleic acid 43 to 45% and oleic acid 31% (Ecky, 1954). Total unsaturated fatty acid composition is 54.53% (Al Ashaal et al., 2010). Moreover, Grant et al. (1990) reported that low dietary linoleic acid intake increases the incidence of duodenal ulcers. It is therefore believed that the fatty acid components of B. aegyptiaca oil used in this work may have contributed to the observ-

ed moderate anti-ulcer property. Other consti-tuents of B. aegyptiaca oil such as flavonoids (anti-oxidants) may also have contributed to its anti-ulcer activity.

The results of the present investigation suggested that consumption of B. aegyptiaca oil would be beneficial for patients suffering peptic ulcer disease. From this report, B. aegyptiaca oil possessed both gastric anti-secretory and gastric cyto-protective effects. However, the effects of B. aegyptiaca oil on the growth of H. pylori, one of the main causes for the development of gastric and duodenal ulcer is not known. Studies on the effect of B. aegyptiaca oil on the H. pylori infection, gastric PH, gastric HCl and total acids have to be carried out to further support its beneficial effect in peptic ulcer patients. A synergistic composition of B. aegyptiaca oil and other anti-ulcer oils such as fish oil (Riber et al., 1999) is also suggested in the prevention, treatment and relief of the symptoms of peptic ulcer disease. REFERENCES Aiello ES, Mays A (1998).The Merck Veterinary Manual. 8

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