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Effects of Ethyl acetate Portion of Syzygium Aromaticum Flower Bud Extract on Indomethacin-Induced Gastric Ulceration and Gastric Secretion 908 30 minutes all the groups were administered indomethacin (20mg/kg S.C.).Then, the animals were deprived of both food and water for seven (7) hours (Urushidani et al., 1979). Later on, the animals were sacrificed by decapitation (Satoh et al., 1983). The stomachs were opened along the greater curvature, rinsed slowly with normal saline then stretched out as much as possible on a Whatman’s filter paper. The ulcerated surface in each stomach was measured with a transparent millimeter scale rule and the result for each group was expressed in mm of mean ulcer ± S.E. (Ulcer index) (Scepovic and Radmanovic, 1984). Preventive index (P.I. %) was later calculated according to the method of Hano et al. (1976) as follows: U.I. indomethacin – U.I. extract plus indomethacin P.I. % = X 100 U.I. indomethacin 2.7. Study of Gastric Secretion Gastric juice was collected after making pyloric ligation according to the technique of Shay et al. (1954) as modified by Levine (1965) where esophageal ligation was avoided. The animals were fasted for 48 hours to ensure complete emptying of the stomach, but allowed water ad libitum. The animals were then weighed at the end of the fasting period. Then, under light anaesthesia, the abdomen was opened by midline incision and the stomach was identified. A pyloric ligature was made by using a thread with care to avoid damage of the blood vessels or traction of the stomach. The abdomen was closed by sutures and cleaned thoroughly with saline. The anaesthesia was discontinued and the animals were allowed to recover for a period of three (3) hours. After the 3 hours, the animals were sacrificed and the abdomen of each animal was opened and the esophagus was ligated, then the stomach was removed and washed with saline and dried. An opening along the greater curvature was made and the gastric contents were drained into a graduated centrifuge tube then centrifuged at 3000 rpm for 10 minutes. 2.8. Analysis of the Gastric Juice 2.8.1. Volume of gastric juice After the centrifugation, the supernatant was measured as volume of the gastric juice. 2.8.2. Determination of the Titritable Acidity A given volume of the gastric juice (0.2 ml) was titrated against 0.01 N NaOH using an end point of pH 7.0 as determined colorimetrically with phenol red as an indicator (Grossman, 1963). It was calculated as milliequivalents per liter (mEq/L) which is equal to the number of milliliters (mL) of 0.1 N NaOH required to neutralize 100 ml of the gastric juice Volume of 0.01N NaOH which neutralizes 1 mL of gastric juice = X 100 10 (Davenport, 1977). 2.8.3. Determination of acid output This was calculated as µEq/h by multiplying the volume of gastric secretion (ml/h) of an animal by the titritable acidity (acid concentration) (mEq/L) of the gastric secretion in that animal (Okabe et al., 1975).
909 Okasha Mohammad Abdul- Halim, Magaji Rabiu Abdussalam, Abubakar Mujtaba Suleiman and Fatihu Muhammad Yakasai 2.9. Statistical Analysis The experimental data were expressed as the mean ± S.E.M. The significance of the results was determined using analysis of variance (ANOVA) with later comparison by Dunnett’s t-test and the results were considered significant if p < 0.05. 3. Results 3.1. Phytochemical Screening Preliminary phytochemical analysis of the extract revealed the presence of flavonoids, glycosides, tannins, resins and alkaloids. 3.2. Acute Toxicity Studies The intraperitoneal LD50 of the extract in mice was found to be 565.7mg/kg. 3.3. Gastric Mucosal Integrity Studies Indomethacin (20 mg/kg S.C.) produced severe gastric mucosal ulceration restricted to the corpus with an ulcer index of 33.9 ± 1.2. The ulcers appeared in the form of streaks and/or groups and went deep in the mucosa with severe necrotic and hemorrhagic bases. Administration of SaETAC (100 mg/kg) showed a significant (p
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