To investigate the regulatory effect of berberine on local AMP-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR) targets of ovaries in rats with polycystic ovary syndrome(PCOS) and thereby to explore the mechanism of berberine in improving local insulin resistance(IR) of ovary.Methods:A total of 60 female Sprague-Dawley(SD) rats were randomized into blank group，model group，berberine group，and metformin group，with 15 rats in each group.The Biosynthetic Human Insulin Injection(NS) and human chorionic gonadotropin(HCG) were used to induce PCOS in rats except for the blank group.Berberine group and metformin group were given(ig) berberine and metformin，separately.After the intervention，serum sex hormone levels and glucose and lipid metabolism were detected in each group，and the morphological changes of ovary were observed.The protein expression of α-pT172-AMPK，pSer2481-mTOR，and pSer473-AKT in ovary was detected by Western Blotting，and the mRNA expression of glucose transporter 4(GLUT4) in ovary by real-time fluorescent quantitative PCR.Results:1) Compared with the blank group，the model group showed significantly high levels of the hormones serum total testosterone(T)，luteinizing hormone(LH)，LH/follicle-stimulating hormone(FSH)，17α-hydroxyprogesterone(17α-OHP)，and androstenedione(ASD)，significantly low level of sex hormone binding globulin(SHBG)，and significantly high levels of fasting insulin(FINS) and homeostasis model assessment of insulin resistance(HOMA-IR).The ovary showed many cystic follicles and reduction in the number of follicles and granulosa cell layers of follicles at each development stage.Moreover，significant decrease in the levels of α-pT172-AMPK and pSer473-AKT and the mRNA expression of GLUT4 and significant increase in the protein level of pSer2481-mTOR in the ovary were observed in the model group as compared with those in the blank group.2) Compared with the model group，berberine group demonstrated significant improvement in serum reproductive metabolism indexes，recovery of ovary morphology，significant increase in the levels of α-pT172-AMPK and pSer473-AKT in ovarian tissues and mRNA expression of GLUT4，and significant reduction in protein level of pSer2481-mTOR.Conclusion:Berberine can alleviate the reproductive-endocrine disorder，regulate IR，and restore the development of ovarian follicles of PCOS rats.The mechanism is the likelihood that berberine acts on AMPK/mTOR and modulates local insulin signal transduction.