ANTIDIABETIC ACTIVITY OF THE FRUITS POWDER OF CISSUS ROTUNDIFOLIA ON ALLOXAN - INDUCED DIABETIC RABBITS

Dr.Wael Mustafa Ali Mohammed. Department of Biology, Faculty of Sciences, Aden university ,Yemen. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History Received: 10 September 2019 Final Accepted: 12 October 2019 Published: November 2019


Experimental animals:
The animals (locally rabbits) were purchased from the local market in Lahi governorate, Yemen. The rabbits were divided randomized into sex groups comprising of 10 animals weighed 1200-1350g in each group. They were allowed to acclimatize for two weeks and were maintained. All animals were given free access to standard chow and tap water. The fruits powder suspension was administered orally twice per day.

Induction of experimental diabetes
Diabetes mellitus was induced in rabbits by single intraperitoneal injection of freshly prepared solution of alloxan (120 mg/kg b.w) in physiological saline after overnight fasting for 12 h 24 .marginal ear vein blood glucose levels were measured, 7 ,15 and 30 days after alloxan injection .The rabbits were fed by the preparations by gastric intubation.

Experimental design
1. Group I: Normal control rabbits received normal saline 10 ml /kg/day and free food for 30 days 2. Group II: Diabetic control rabbits received alloxan in single dose (120 mg/kg, i.p.).did not any treat. 3. Group III: Treated diabetic rabbits received CRFP (1g /kg/body weight, in 2 ml distilled water daily, for 30 days. 4. Group IV: Treated diabetic rabbits received CRFP (2g/kg/ body weight, in 2 ml distilled water daily, for 30 days. 5. Group V: Treated diabetic rabbits received CRFP (3g /kg/ body weight, in 2 ml distilled water daily, for 30 days. 6. Group VI: Diabetic rabbits treated with glibenclamide (5 mg/kg/ body weight, in 2 ml distilled water daily, for 30 days).

Statistical analysis:-
The results are expressed by the means and standard deviations(SD). The statistical analysis was carried out using paired t-test and One-way analysis (ANOVA). Statistical P. value at the level of <0.05 was considered to be significant.

Results:-
The oral administration of CRFP of Cissus rotundifolia leaves didn't cause any signs of clinical abnormalities in the treated rabbits. Thus the drug dose given appears to be safe. Continuous treatment with CRFP (1, 2 and 3g/kg) for a period of 30 days showed a significant decrease in the blood glucose level in diabetic rabbits. A maximum reduction of blood glucose level was occurred at the dose of 3 g/kg. p. o. of CRFP . Table NO (1).

Discussion:-
NO lethality or any toxic reactions were found at any of the selected CRFP doses until the end of the study period. The oral administration of CRFP of Cissus rotundifolia fruits didn't cause any signs of clinical abnormalities in the treated rabbits. Thus the CRFP dose given appears to be safe. Toxicity Studies did not show any side effects of cissus genus 25 . Alloxan has been observed to cause a massive reduction of the β-cells of the islets of Langerhans and induce hyperglycemic 26,27 . The mechanism behind alloxan diabetogenic effect is not very clear. However, it has been suggested that it damages pancreatic beta cells through formation of superoxide anions and hydrogen peroxide but not hydroxyl radicals 28,29 . This causes DNA strand-breaks (30) and depletes NAD+ stores 31,32 . One study suggested that the generation of free radicals through redox cycling with glutathione was not enough to cause diabetes by alloxan ,but it seems that hydrophilicity of this compound is a necessary condition for its diabetogenecity 33 . Some reports indicate that the formation of superoxide and hydroxyl radicals is responsible for the alloxan-induced cytotoxicity 34,35,36 . Pancreas from alloxan-induced diabetic animals showed a 70% reduction of beta cells areas. At the same time, there was an increase in the areas occupied by delta cells, and no alteration in glucagon-producing cells 37 .

Mechanism of action of the standard drug glibenclamide:
Binding of glibenclamide with its receptor leads to the closure of the potassium channels which opens calcium channels for influx of Ca2+ ions into the cytoplasm and release of insulin from the pancreatic islets. These K+ channels are responsive to ATP/ADP ratio and close when the ratio increases because of an increase in glucose metabolism 38,39 . Sulphonylureas such as glibenclamide stimulate insulin secretion from pancreatic β cells principally by inhibiting ATP-sensitive K+ channels 40 .
Possible Mechanism of action of the phinolic compounds found in Cissus rotundifolia: This study showed that the CRFP of Cissus Rotundifolia cause a significant hypoglycemic effect in alloxan diabetic rabbits. The maximum reduction in fasting blood glucose (FBG) was observed at a dose of 3 g /kg b.w. Thus, g /kg/.w. was found to be the optimum dose of CRFP on FBG of diabetic animals. These findings are supported by those of 21.22.23 . There were a lot of studies that support the anti-hyperglycemic or anti-diabetic effect of several medicinal plants worldwide 41,42,43 . A number of researchers have reported that genus cissus have antidiabetic properties in various studies 44,45,46,47,48,49,50 . The cissus genus (vitaceae) extracts are characterized by a high content of flavonoids. An important source of flavonoids in the diet and the flavonoids found in cissus genus are known to be strong antioxidants 51 . Conclusion: The antihyperglycaemic activity may be due to the presence of several bioactive anti-diabetic principals.