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Journal of Medicinal Plants Research Vol. 6(12), pp. 2266-2275, 30 March, 2012 Available online at http://www.academicjournals.org/JMPR DOI: 10.5897/JMPR10.409 ISSN 1996-0875 ©2012 <strong>Academic</strong> <strong>Journals</strong> Full Length Research Paper Anti-ulcer activity of Swietenia mahagoni leaf extract in ethanol-induced gastric mucosal damage in rats Salmah Al-Radahe 1 , Khaled Abdul-Aziz Ahmed 2,3 *, Suzy Salama 2 , Mahmood Ameen Abdulla 2 , Zahra A. Amin 2 , Saad Al-Jassabi 1 and Harita Hashim 4 1 Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia. 2 Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. 3 Department of Medical Sciences, Faculty of Dentistry, Ibb University, P. O. Box 70627, Ibb, Yemen. 4 Department of Biology, Faculty of Applied Science, University Teknologi MARA 40450 Shah Alam, Malaysia. Accepted 13 April, 2011 Swietenia mahagoni (West Indian mahogany) has been reported to have medicinal uses, such as treatment for hypertension, cancer, amoebiasis, chest pains and intestinal parasitism. The present study was performed to evaluate the acute toxicity and anti-ulcer activity of S. mahagoni ethanol leaf extract against ethanol-induced gastric ulcer. 24 rats included in this study were divided into 4 groups with 6 rats each group. Group 1 rats (ulcer control group) were pre-treated with vehicle (Carboxyl methyl cellulose). Group 2 (reference group) was orally pretreated with 20 mg/kg omeprazole. Group 3 and 4 (experimental groups) were orally pre-treated with S. mahagoni ethanol leaf extract at 250 and 500 mg/kg doses, respectively. After one hour later, all groups received absolute ethanol to generate gastric mucosal injury. After an additional hour, all the rats were sacrificed and the ulcer areas of the gastric walls were determined. Grossly, the ulcer control group exhibited severe mucosal injury, whereas pretreatment with either omeprazole or plant extract resulted in significantly protection of gastric mucosal injury and increase in mucus production. Flattening of gastric mucosal folds was also observed in rats pretreated with S. mahagoni leaf extract. Histological studies of the gastric wall of ulcer control group revealed severe damage of gastric mucosa, along with edema and leucocytes infiltration of the submucosal layer compared to rats received either omeprazole or S. mahagoni leaf extract where there was marked gastric protection along with reduction or absence of edema and leucocytes infiltration of the submucosal layer. Acute toxicity study with a higher dose of plant extract at 5 g/kg did not reveal any toxicological signs in rats. In conclusions, the present findings suggest that S. mahagoni ethanol leaf extract exhibit an anti-ulcer activity against ethanol-induced gastric ulcer in experimental animals. Key words: Swietenia mahagoni leaf extract, gastric ulcer, histology. INTRODUCTION Various parts of Swietenia mahagoni have been used as folk medicine for the treatment of hypertension, malaria, cancer, amoebiasis, chest pains, fever, anemia diarrhea, dysentery, depurative and intestinal parasitism (Nagalakshmi et al., 2001; Maiti et al., 2007). Other studies have showed other medicinal values of S. mahagoni plant like antibacterial (Majid et al., 2004) and *Corresponding author. E-mail: khaaah@gmail.com. antioxidant activities (Sahgal et al., 2009a) as well as diabetes therapy (Li et al., 2005). The biologically active ingredients, tetranortriterpenoids and fatty acids, are considered to be responsible for these therapeutic effects (Bacsal et al., 1997). Previous phytochmical investigations on S. mahogani have led to the isolation of more than 45 limonoids belonging to the structural types of andirobin, gendunin, mexicanolide, phragmalin, triterpens, tetranortriterpenes, and chlorgenic acid (Abdelgaleil et al., 2006; Chen et al., 2007). From S. Mahagoni, eighteen tetranotriterpenoids Wer isolated (Kadota et al., 1990) and the presence of
known fatty acids and terpenoids were reported (Saad et al., 2003). There are no data available regarding antiulcerogenic property of S. mahagoni in rats. Therefore, the present study was undertaken to evaluate the antiulcerogenic activity of S. mahagoni ethanol leaf extract against ethanol-induced gastric mucosal damage in experimental rats. MATERIALS AND METHODS Omeprazole Omeprazole, a proton pump inhibitor, has been widely used as an acid inhibitor agent for the treatment of disorders related to gastric acid secretion for about 15 years (Li et al., 2004). In this study, Omeprazole was used as the reference anti-ulcer drug, and was obtained from the University of Malaya Medical Centre (UMMC) Pharmacy. The drug was dissolved in carboxylmethyl cellulose (CMC) and administered orally to the rats in concentrations of 20 mg/kg body weight (5 ml/kg) (Pedernera et al., 2006). Plant material S. mahagoni leaves were obtained from Ethno Resources Company (Selangor Malaysia) and identified by comparison with the Voucher specimen deposited at the Herbarium of Rimba Ilmu, Institute of Biological Sciences, University of Malaya, Malaysia. Preparation of plant extract S. mahagoni leaves were shade-dried for 7 to 10 days and were then powdered using electrical blender. 100 g of fine powder were soaked in 500 ml of 95% ethanol in conical flask for 3 days. After 3 days the mixture was filtered using a fine muslin cloth followed by paper filtration (Whatman No. 1) and distilled under reduced pressure in an Eyela rotary evaporator (Sigma-Aldrich, USA). The dry extract was then dissolved in CMC (0.25% w/v) and administered orally to rats in concentrations of 250 and 500 mg/kg body weight (5 ml/kg body weight) (De Pasquale et al., 1995). Acute toxicity test Experimental animals Adult healthy male and female Sprague Dawley rats (6 to 8 weeks old) were obtained from the Animal House, Faculty of Medicine, University of Malaya, Kuala Lumpur. The rats weighed between 150 to 180 g. The animals were given standard pellets and tap water. The acute toxicity study was used to determine the safe dose for the plant extract. Thirty six Sprague Dawley rats (18 males and 18 females) were assigned equally into 3 groups. The first group was labeled as vehicle (CMC, 0.25% w/v, 5 ml/kg) while the second and third groups of animals were pretreated with 2 and 5 g/kg of S. mahagoni leaf extract, respectively. The animals were fasted overnight (food but not water) prior dosing. Food was withheld for a further 3 to 4 h after dosing. The animals were observed for 30 min and 2, 4, 8, 24 and 48 h after extract administration for the onset of clinical and/or toxicological symptoms. The animals were sacrificed on the 14 th day and histological, hematological and serum biochemical parameters were Al-Radahe et al. 2267 determined following standard methods (Bergmeyer and Horder, 1980; Tietz et al., 1983). Behavioral observation and mortality Throughout the study period, all animals were observed for behavioral signs of toxicity, morbidity and mortality. Mortality checks were made twice daily and determination of behavioral signs was observed daily for all animals. Detailed observations of the individual animals were made weekly in comparison with the vehicle treated animals. Observations included gross evaluations of the skin, any signs of respiration (dyspnea), salivation, exophthalmia, convulsion and any changes in locomotion such as whether the animals tend to stay quietly or actively moving in their cage. Hematological and biochemistry analysis The animals were fasted overnight prior to necropsy and blood was collected. Blood samples were drawn from jugular vein under diethyl ether anesthesia. Blood samples were collected into EDTA tubes for total and differential white blood cell (WBC) count. For serum biochemistry analysis, blood was collected into anticoagulant-free tubes. Biochemical parameters include aspartate aminotransferase (AST), alanine aminotransferase (ALT), total protein, albumin, globulin, total bilirubin, conjugated bilirubin, alkaline phosphatase, gamma glutamyl transferase (GGT), urea, creatinine, anion gap and serum electrolytes (CO2, Potassium, Sodium and Chloride). All samples were sent immediately to the Clinical Diagnostic Laboratory at University of Malaya Medical Centre for liver and renal function tests. The results were compared to that of the rats’ respective control groups. Gross necropsy and histopathology At scheduled termination, all surviving animals were anesthetized by diethyl ether inhalation and quickly sacrificed by exsanguinations of jugular vein for blood sample collection. Gross postmortem examinations were performed on all terminated animals. Liver and kidney from each animal were routinely processed and embedded in paraffin. After sectioning and staining with Haematoxylin and Eosin (H and E) stain method, all slides were observed under microscope to observe for any pathological changes. Anti-ulcer activity studies Experimental animals Sprague Dawley healthy adult male rats were obtained from the Experimental Animal House, Faculty of Medicine, University of Malaya. The animals were kept at room temperature in humidity rooms on a standard light/dark cycle (12 h light; 12 h dark cycle). The rats were divided randomly into 4 groups of 6 rats each. Each rat that weighted between 200 to 225 g was placed individually in separate cage (one rat per cage) with wide-mesh wire bottoms to prevent coprophagia during the experiment. The animals were maintained on standard pellet diet and tap water. Gastric ulcer induction by ethanol and tissue sample collection The rats were fasted for 48 h before the experiment (Garg et al.,
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ACKNOWLEDGMENTS The authors would l
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Figure 3. The most widely included
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et al., 1996; van Wyk and Gericke,
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DPPH inhibition (%) DPPH % inhibiti
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% Mortality Concentration (µg/ml)
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information on plants used for the
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Warghat et al. 2391 Table 3. Summar
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Excoffier L, Smouse PE, Quattro JM
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DPPH• scavenging ratio(%) OD valu
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scavenging effect on free radical a
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our new CL system. Fenton reaction
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1 2 Time (s) Figure 2. Kinetic curv
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Pan YM, Liang Y, Wang HS, Liang M (
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UPCOMING CONFERENCES 15th European
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