A multifactorial endocrinopathy affecting women in their reproductive years, polycystic ovarian syndrome (PCOS) causes insulin resistance (IR), hyperandrogenism, cardiovascular issues, obesity, and menstruation difficulties. The present study was designed to investigate the therapeutic potentials of the methanolic extract of Iris ensata and standard drug metformin (MET) on letrozole (1 mg/kg) induced against estrus cyclicity, biochemical and hormonal aspects of polycystic ovary syndrome in Wistar rats. Twenty-four Wistar rats were divided into four groups including a plain control group, a negative control group (PCOS), a positive control group and a treatment group. The positive control group received MET (150 mg/kg body weight) for 21 days. PCOS was induced with letrozole (1 mg/kg body weight) for 21 days. The treatment group was treated with a methanolic extract of Iris ensata for three weeks after the induction of PCOS for 21 days. Body weight and estrous cycle phase were measured every day. Rats were sacrificed after 21 days of treatment. Blood samples were collected on 14, 21 and 42 days for the measurement of biochemical profile i.e. glucose, triglycerides, total cholesterol and hormone levels i.e. testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estradiol. There was letrozole-induced PCOS characterized by irregular estrus cyclicity, elevated triglycerides, total cholesterol, luteinizing hormone (LH), testosterone and estradiol concentrations compared with the control. These reproductive, biochemical, and hormonal alterations were alleviated by Iris ensata extracts. For instance, Iris ensata restored the estrus cyclicity with a remarkable effect after 21 days of treatment. Moreover, Iris ensata significantly decreased (p<0.001) LH and testosterone levels, but increased (p<0.01) estradiol (methanolic extract; 125 mg/kg) concentration. Iris ensata alleviated reproductive, biochemical and hormonal alterations in PCOS rats and could be useful in the management/treatment of reproductive and metabolic disorders related to PCOS.