Therapeutic potentials of the caffeine in polycystic ovary syndrome in a rat model: via modulation of proinflammatory cytokines and antioxidant activity Running title: The effects of caffeine on rats with polycystic ovary syndrome

Recent studies have shown that 6-20% of women worldwide suffer from Polycystic ovary syndrome.It is associated with reproductive and metabolic dysfunction.Caffeine is naturally found in tea, cocoa, and coffee.It has shown that caffeine can change hormonal profiles, stimulate ovulation, and enhancing fertility. In this study,the effects of caffeine on PCOS rats were investigated. For this purpose, forty female rats were divided into four groups: the control group(without injection),the PCOS group(injection of 2 mg of estradiol valerate to each rat), the Caffeine group(37.5 mg/kg caffeine),and the PCOS + caffeine group(37.5 mg/kg caffeine).After 21 days of treatment, the ovaries of rats were removed and prepared for further evaluation including H&E staining,TUNEL, Real-time PCR, and biochemical analysis.Administration of caffeine in PCOS mice considerably reduced both the volume of the ovary(p<0.05)and follicular clusters(P<0.01).However, SOD and GPX enzymes were dramatically active in the PCOS + caffeine group compared to the rest(P<0.05). Besides, caffeine treatment in PCOS mice led to Bax reduction and increase of Bcl2 expression. On the other hand, the expression of IL-6 and TNF in caffeine + PCOS were high compared to the other groups.We found that caffeine can reduce apoptosis and inflammation in PCOS ovaries and enhance the unpleasant symptoms of PCOS