高雄激素影响卵泡发育过程中甲基化/去甲基化稳态的分子调节机制

Hyperandrogenism is the most important characteristics of PCOS, especially in the follicular fluid. Also, hyperandrogenism is the basic pathogenesis for ovarian dysfunction (without dominant follicle development or ovulation). In recent years, epigenetic regulation has been reported to play a role during follicular development. Our studies showed that androgen can downregulated the expression of DNMT3a and upregualate imprinted gene expression in human granulosa cells （GCs）, suggesting that androgen may regulate the meytlation level during follicular development. Based on these, we will perform whole genome DNA methylation sequencing and pyrosequencing on oocyte from androgen treated mosue to check methylation level and DMR. With in vitro androgen stimulation and SiRNA knockdown on human GCs, we will determine the key enzymes which involved in methyltion or demethylation. Finally, we will investigate the mechanism that androgen regulated the target enzymes by ChIP-assay and luciferase reporter. Therefor, from the point of metylation/demethylation, the current project will provide theory basis for clinical prediction and intervention of PCOS.

卵泡局部高雄激素水平是多囊卵巢综合征(PCOS)的重要生化特征，也是导致卵泡发育停滞，排卵障碍的主要病理生理基础。近年，表观遗传学研究已经逐渐成为调节卵泡发育和PCOS的焦点。本实验室的前期研究发现，高浓度雄激素可下调人颗粒细胞中甲基化转移酶DNMT3a的表达，同时伴有母源性印记基因甲基化水平的降低及表达升高，提示高雄激素可能影响卵泡发育过程中的甲基化水平。本项目拟通过全基因组甲基化测序观察高雄大鼠卵母细胞/颗粒细胞整体甲基化水平改变，利用焦磷酸盐测序验证差异甲基化去（DMR）甲基化状态，对体外培养颗粒细胞（GCs）进行siRNA干扰，筛选雄激素作用的甲基化/去甲基化通路关键酶，最后通过ChIP、荧光素酶报告等手段阐明其分子机制，进而从甲基化/去甲基化稳态调节的表观遗传学层面，揭示高雄激素影响卵泡发育的机制，为PCOS的临床治疗提供理论依据。