The Effect And Mechanism Of Cryptotanshinone On Steroidogenesis And Glucose Metabolism In PCOS Ovarian Granulosa Cells

Objective:The present study is to evaluate the ability of steroidogenesis and glucose metabolism in Polycystic ovary syndrome(PCOS)ovarian granulosa cells in vitro,and to investigate the effects and molecular mechanism of cryptotanshinone on PCOS granulosa cells steroidogenesis and glucose metabolism.Methods:The patients with or without PCOS were recruited from women undergoing in vitro fertilization/Intracytoplasmic sperm injection-embryo transfer(IVF/ICSI-ET)according to the inclusion and exclusion criteria.Follicular aspirate was collected from the subjects,then the ovarian granulosa cells were isolated and cultured in vitro.The PCOS ovarian granulosa cells were treated with cryptotashinone or AICAR,an activator of AMP-activated protein kinase(AMPK)for 48h.Three expetiments were performed.The level of progesterone,testosterone and estradiol in cell cultrue media were measured by chemiluminescent immunoassay.Glucose assay kit was performed to determine the glucose in cultured medium.Real-Time Quantitative PCR was performed to detect the expressions of AMPK,Steroidogenic acute regulatory protein(StAR),3β-hydroxysteroid dehydrogenase(HSD3p),17-Hydroxylase/C 17,20-lyase(CYP17),Cytochrome P450(CYP19),Glucose transporter 4(GLUT4).Results:Compared with the control group,the concentration of testosterone in the culture medium of PCOS ovarian granulosa cells was obviously increased,and the progesterone and estradiol levels were obviously decreased.The glucose level in the culture medium was significantly elevated after insulin stimulation.The expressions of StAR,3βHSD and CYP17 mRNA were elevated,while the levels of AMPK and CYP19 mRNA were decreased as well as GLUT4 gene expression.Compared with PCOS granulosa cells group,Cryptotanshinone treatment decreased the level of testosterone and increased the levels of progesterone and estradiol.Glucose level in culture medium was decreased.The levels of StAR,HSD3β and CYP17 expression were decreased,while AMPK,CYP19 and GLUT4 gene expressions were all increased.Above-mentioned genes expression levels in PCOS granulosa cells with AICAR treatment were consistent to the results of cryptotanshinone treatment group.Conclusions:(1)Abnormal glucose metabolism and steroidogenic production were preseneed in the ovarian granulosa cells in PCOS patients.(2)Abnormal steroid production in ovarian cells was involved in ovary dysfunction in women with PCOS.(3)Inhibition of androgen production by cryptotanshinone was mediated by steroidogenic enzymes.(4)AMPK pathway could be the mechanism of improvement of abnormal glucose metabolism and excess androgen in PCOS.