ISPUB.COM The Internet Journal of Pharmacology Volume 7 Number 1 Anti-ulcer effect of Cordia dichotoma Forst.f. fruits against gastric ulcers in rats. I Kuppast, P Vasudeva Nayak, K Chandra Prakash, K Satsh Kumar Citation I Kuppast, P Vasudeva Nayak, K Chandra Prakash, K Satsh Kumar. Anti-ulcer effect of Cordia dichotoma Forst.f. fruits against gastric ulcers in rats.. The Internet Journal of Pharmacology. 2008 Volume 7 Number 1. Abstract The anti-ulcer effect of extracts of Cordia dichotoma Forst.f. fruits (300mg/kg body weight) was studied in albino rats of Wistar strain using three different models i.e. pyloric ligation, aspirin and indomethacin induced ulcers. The extractions of C.dichotoma Forst.f. fruits were carried out using ethanol. This extract was fractionated using petroleum ether, solvent ether, ethyl acetate, butanol and butanone in succession. Gastric mucosal injury was produced in rats by pyloric ligation, aspirin and indomethacin induced models. Extracts of petroleum ether, solvent ether, ethyl acetate, butanol and butanone were administered in a dose of 300 mg/kg body weight. The parameters taken to assess anti-ulcer activity were volume of gastric secretion, free acidity, total acidity and ulcer index. The results indicates that, extracts of ethyl acetate, butanol and butanone significantly (p< 0.001) decrease the volume of gastric secretion, free acidity, total acidity and ulcer index with respect to control. The results suggest that the extracts of Cordia dichotoma Forst.f. fruits possess significant anti-ulcer activity. INTRODUCTION >C. dichotoma Forst.f. a plant belonging to family Boraginaceae is medium sized tree with a short, usually crooked trunk 3-4 ft. in girth 1. The fruits are globose, yellowish-brown, pink or black and pulpy. The plant grows in India, Sri Lanka and other warmer regions. The medicinal attributes of C.dichotoma have been known since a long time. The fruits of the plant are used as cooling, astringent, emollient, expectorant, anthelmintic, purgative and diuretic 2. A number of pharmacological properties such as analgesic, antiinflammatory and hepatoprotective have been reported 345. The study assumes significance in the context that prolonged use of synthetic anti-ulcer drugs leads to adverse drug reactions and search for new anti-ulcer agents that retain therapeutic efficacy and are devoid of adverse drug reaction is warranted. A study of the efficacy of an extracts of C. dichotoma fruits in gastric ulcer with pyloric ligation, aspirin and indomethacin induced ulcers was under taken in rat models. College, Hubli. The collected fruits were shade dried, reduced to coarse powder (2 kg) and subjected to repeated exhaustive extraction in batches with ethanol in a soxhlet extractor. After complete extraction the alcoholic extract (68 gm) was then suspended in water and further fractionated using petroleum ether (8 gm), solvent ether (2 gm), ethyl acetate (4 gm) butanol (5.5 gm) and butanon (2.5) in succession. These extracts were vacuum dried and used for anti-ulcer activity. Tween 80 (1%) was used as vehicle to suspend the extracts. PHYTOCHEMICAL SCREENING All the extracts were subjected to phytochemical analysis as per standard procedures 6 to know the nature of phytoconstituents present (TABLE 1). The major active constituents were found in extracts includes flavanoids, alkaloids and saponins. Among all active constituents flavanoids were found to be in higher concentrations. MATERIALS AND METHODS PREPARATION OF PLANT EXTRACTS The fruits of C.dichotoma were purchased from local market of Hubli and were authenticated by Prof V. S Huddar, Department of Botany, H. S. K Science and S. K Arts 1 of 5
Figure 1 TABLE 1: Qualitative chemical examination of various extracts obtained by successive solvent fractionation of the. fruits. dose of 300mg/kg body weight for 8 days p.o. After 8 days of treatment, the animals were fasted for 24 hours. Ulcer was produced by administration of aspirin (a dose of 200 mg/ kg orally) on the day of sacrifice. The animals were sacrificed 4 hour later and stomach was opened to calculate the ulcer index by Kunchandy method 8. INDOMETHACIN INDUCED ULCER MODEL Indomethacin was administered orally in a dose of 15 mg/kg 9 body weight. The animals were sacrificed 6 hours after Indomethacin administration. ANIMALS The experiments were initiated only after approval Institutional Animal Ethical Committee. Albino mice weighing 25-30 g and albino rats weighing 150-200 gm were obtained from central animal house, K. L. E. S s College of Pharmacy, Hubli, Karnataka. They were maintained at standard housing conditions and fed with commercial diet (Hindustan Lever Ltd., Bangalore) and water ad libitum during the experiment. Figure 2 Table 2: Effect of fruits extracts and Ranitidine in pyloric ligated in rats ACUTE TOXICITY STUDIES The acute toxicity studies were carried out as per stair case or Up and down method 7. Accordingly the LD 50 of all extracts were found to be 3000 mg/kg bodyweight. One tenth of this dose was selected for the study of aphrodisiac activity. Figure 3 Table 3: Effect of fruits extracts and ranitidine in aspirin induced ulcer in rats EXPERIMENTAL DESIGN FOR ANTI-ULCER ACTIVITY The Albino rats of Wistar strain are divided into seven groups of six animals each. Animals were fasted 24 hours. First group received normal saline 2ml/kg body weight (negative control), the second group received ranitidine 20mg/kg body weight by oral route (positive group) and reaming group III, IV, V, VI and VII were received extracts of petroleum ether, solvent ether, ethyl acetate, butanol and butanone respectively in the dose of 300mg/kg body weight by oral route. PYLORIC LIGATION METHOD In this method extracts were administered 30 minutes prior to pyloric ligation, Animals were scarified 4 hour later and the stomach was opened to collect the gastric contents. The total volume of gastric content was measured. ASPIRIN INDUCED ULCER MODEL In this method the treated groups received extracts in the 2 of 5
Figure 4 Table 4: Effect of fruits extracts and ranitidine in indomethacin induced ulcer in rats All values are in mean ± SE. n=6, VGJ = Volume of gastric juice in ml, TA = Total acidity in meq/l., FA = Free acidity in meq/l. UI = Ulcer index, P <0.001 = *, The volume of Sodium hydroxide required corresponds to the total acidity. Acidity (meq /l/100g) was calculated as; Acidity = {Volume of NaOH X Normality X 100 meq/l/100g X 0.1} MACROSCOPIC EXAMINATION The stomach was cut open along the greater curvature and washed in Normal saline. The gastric juice was collected in a measuring cylinder. The gastric juices were centrifuged at 1000 rpm for 10 min. 1 ml of the supernatant was diluted with 9 ml of distilled water. The solution was titrated against 0.1 N sodium hydroxide using Topferu reagent till the solution turned to orange colour. Then it was laid flat and the ulcer area was calculated under a dissecting microscope (10xs) with a square grid. Gastric mucosal lesions were seen in the form of haemorrhage or linear breaks. Ulcer index was calculated using the following method Ulcer index = 10/x Where x = Total mucosal area / Total area of mucosal lesions The effect of extracts of C. dichotoma fruits on pyloric ligation, aspirin and indomethacin induced ulcer model is presented in Table 2, 3 and 4 respectively. STATISTICAL ANALYSIS All the results were reported as mean S.E. Significance of the difference between control and drug treated were determined by the student t test RESULTS The LD 50 of the extracts were found to be 3000mg/kg body weight by Up and Down method and 1/10 th of the LD 50 i.e. 300mg/kg body weight was considered as therapeutic dose. All the results were analysed by student t test and level of significance was P<0.001. In the present study a dose of 300 mg was found to protect gastric mucosa from ulcer. Therefore, we have used the same dose in this study. The effect of fruit extracts of C. dichotoma on pyloric ligated rat, aspirin and indomethacin induced ulcer models are presented in 1, 2 and 3 respectively. The results of the present study indicates that the ethyl acetate butanol and butanone extracts significantly reduces the total volume of gastric juice, free and total acidity of gastric secretion against and also has activity against gastric ulcer in rats. The control animals had ulcers and haemorrhagic streaks, where as in animals administered with the ethyl acetate, butanol and butanone extracts of C.dichotoma fruits there was significant reduction in ulcer index (P< 0.001) (Figure-1). The results are summarised in Table 1, 2, and 3. However ranitidine prevented ulceration completely. The present study confirms our preliminary observations that Fruit of C. dichotoma Forst.f. have anti-ulcer activity. DISCUSSION It is generally accepted that gastric ulcers results from an imbalance between aggressive factors and the maintenance of the mucosal integrity through endogenous defence mechanisms 9. The excess gastric acid formation by prostaglandins (PG) includes both increases in mucosal resistance as well as a decrease in aggressive factors, mainly acid and pepsin 10. Inhibition of PG synthesis by aspirin coincides with the earlier stages of damage to the cell membrane of mucosa, parietal and endothelial cells 11. The drug reported to be anti-ulcer in traditional system of medicine and used for that purpose has been shown in rat s significantly reduced ulcer activity. The preliminary phytochemical studies revealed the presence of flavanoids in all above three extracts of C. dichotoma; various flavanoids have been reported for its anti-ulcerogenic activity with good level of gastric protection 12. So the possible mechanism of anti-ulcer action of C. dichotoma fruits may be due to its flavanoids content. In this study we observed that C. dichotoma provides significant anti-ulcer and cytoprotective effect against gastric ulcers in rats. Further study regarding isolation and characterization of active principal responsible 3 of 5
for anti-ulcer activity is currently under progress. References 1. The wealth of India, Vol. II, A Dictionary of Indian Raw materials and Industrial product; CSIR New Delhi, 1950, 346. 2. Yoganarsimhan S. N., Medicinal Plants of India, Vol. 1, (Interline publishing Pvt. Ltd. Ban galore, Karnataka), 2000, 38. 3. Rapisarda A., Ficarra R., Tommasin S., Caldbro M. L. and Hungsa S., Cordia francisci, C. martinicensis, C. myxa, C. serratifolia and Culmfolia leaves as new source of routine; Analgesic and anti-inflammatory activity, Plant Medica, 1992, 42, 643. 4. Wassel G., El-Menshaw B., Saud A., Meharuna G. and El-Merzabani M., Screening of selected plant for Pyrrolizidine alkaloids and antitumor activity, Pharmazine, 1987, 42, 709. 5. Rajesh M. G, Paul B. and Latha M. S., Efficacy of kamilari in alcoholic liver cirrhosis, Antiseptic., 2000, 97, 320. 6. Evans W. C., Trease G. E., Text book of Pharmacognosy, 13th edition, ELBS, London, 1984. 7. Biggers J. D. and Claringbold P. J., Aust. J. Expti. Biol. Med. Sci., 1954,32, 33. 8. Ghosh M. N., Fundamental of Experimental Pharmacology, 3rd edition, Scientific book Agency, Calcutta, 1986,156. 9. Salma, R. B., Planta Med., 1973, 24, 375 377. 10. Rainsford, K. D., Adv. Inflamm. Res., 1984, 6, 51 64. 11. Alarcon, D. L., Martin, M. J. and Motilva, V., J. Ethnopharmacol., 1994, 42, 161 170. 12. Parmar, N. S. and Shikha Parmar, Indian J. Physiol. Pharmacol., 1998, 48, 343 351. 4 of 5
Author Information I.J. Kuppast P. Vasudeva Nayak Department of Studies in Chemistry, Kuvempu University K. Chandra Prakash K.V. Satsh Kumar 5 of 5