This study aimed to investigate the therapeutic effect of Medical Ozone(MO) in combination with insulin on reproductive hormones and histopathological changes in ovaries of rats with diabetes mellitus (DM). forty eight adult female rats were randomly divided into the following six groups (n = 8): control (A), MO (B), diabetic (C), MOtreated diabetic (E), Insulin-treated diabetic ) E), and MO, Insulin-treated diabetic (F). DM was induced by a single intraperitoneal (ip) Alloxan injection (150 mg/kg ( .after six weeks of treatment Blood sample were collected to estimate biochemical parameters glucose, serum hormones. Also, ovary samples were obtained for histological examination in diabetic untreated rats the results showed hyperglycaemia, and significant decrease in hormones profiles. While ,MO and Insulin treated rats reported an amelioration of the most toxic effect of alloxan and returned most of these parameters nearly normal. Microscopically ovaries showed definite, degeneration in the diabetic group while when use of MO and Insulin treatment in this study showed significant improves of such histological changes when compared to diabetic untreated rats. Conclusion: it was recommended that the use of the Medical Ozone with Insulin as a supplementary agent to reduce oxidative stress damage of hyperglycaemia and recommended to use variable doses and different periods of treatment to evaluate the best dose and period