Preclinical Evaluation of Azadirachta Indica for Treatment of Polycystic Ovarian Syndrome with respect to PI3 Kinase Inhibition

Abstract

Aim and Objective: Polycystic ovarian syndrome is the most common endocrine disorder which affects the women at the reproductive age. A main underlying mechanism associated with PCOS is excessive androgenesis. The inhibition of PI3K in ovary leads to inhibition of 17- hydroxylase enzyme by decreasing the expression of CYP17A1 gene which is associated with hyperandrogenemia. Quercetin has been shown to have beneficial role in treatment of PCOS by inhibition of PI3Kinase. It is also reported that quercetin is abundantly present in leaves of Azadirachta indica. Therefore, the objective of our study is preclinical evaluation of Azadirachta indica in animal model of testosterone propionate induced PCOS with respect to PI-3 kinase inhibition. Materials and methods: Pre pubertal Sprague Dawley (3 weeks old) rats were randomly divided into five groups (n=7): normal control, disease (PCOS) control, diseased animals treated with Azaridichta indica extract (100 mg/kg, p.o.), diseased animals treated with quercetin (100 mg/kg, p.o.) and diseased animals treated with wartmannin (100 mg/kg, p.o.). The PCOS was induced by subcutaneous injection of testosterone propionate (0.2 mg/kg, s.c.) dissolved in olive oil daily for 6 weeks. The animals were examined for the development of PCOS at the end of 6 weeks by estimation of insulin, testosterone and LH from blood. Treatment was continued up to 11 weeks. At the end of the treatment, blood was collected for biochemical estimations and animals were subjected to physical and histopathological evaluations of ovary for total follicular count and for uterus corpus luteum count. Gene expression study of PI3K was also carried out by Polymerase Chain Reaction. Results: The TLC result confirms that quercetin is present in Azadirachta indica extract. In present study, body weight, ovaries weight and uterus weight were found to be decreased by the treatments with Azadirachta indica, quercetin and wartmannin. It was observed that serum glucose, total cholesterol, triglyceride, LDL, VLDL, insulin, testosterone and LH were significantly increased in the diseases control group as compared to normal control group. Treatment with Azadirachta indica, quercetin and wartmannin produced statistically significant decrease in all these parameters. While, the serum level of HDL decreased significantly (P<0.01) in the diseased state and by the treatment with Azadirachta indica HDL levels were found to be improved. Histological examination of ovary and uterus confirmed the disease occurrence and remission state in the diseased treated groups, The PI3kinase gene expression was found to be increased in the diseased group and treatments showed decrease in the expression of PI3kinase gene. Conclusion:The study data suggest that methanolic extract of Azadirachta indica have beneficial effect in PCOS by inhibition of PI3Kinase gene expression and it is also evident from serum biochemical parameters, hormonal parameters. The histopathological exxamination confirm benificial role of Azadirachta indica and quercetin in treatment of PCOS. Beneficial role of Azadirachta indica and quercetin is also supported by gene expression study evocating decrease expression of PI3kinase