miR-93在多囊卵巢综合征卵巢局部中的作用及其机制研究

Ovarian dysfunction and insulin resistance are the key features of polycystic ovary syndrome (PCOS). Studies have confirmed that insulin can regulate the function of ovary in vivo and vitro. Our previous study demonstrated that microRNAs expression altered in the PCOS patients with insulin resistance, and expression of miR-93 was one of the most up-regulate microRNAs in PCOS. MiR-93 promoted proliferation and cell cycle progression of granulosa cells by targeting CDKN1A protein. Furthermore, we found insulin could upregulate the expression of miR-93 and stimulate cell proliferation. In order to clarify the role and molecular mechanism of miR-93 in the pathophysiology of PCOS, we plan to explore the transcription factors and their functinoal roles in differential microRNA expression. By using oligonucleotide array-based transcription factor assay, electrophoretic mobility shift assay, chromatin Immunoprecipitation, and bisulfite sequencing PCR, we will shed light on the role of microRNA participating in insulin regulating the function of ovary, and will provide a new insight of pathophysiology in PCOS and new target for therapies.

卵巢功能障碍、胰岛素抵抗是多囊卵巢综合征（PCOS）重要的病理特征之一，胰岛素参与了卵巢功能的调控，但具体机制不明确。本课题组前期研究证实胰岛素抵抗状态下microRNA是调控卵巢功能的关键分子。我们前期筛选和验证了PCOS卵巢局部miR-93高表达，证实miR-93通过其靶基因细胞周期相关基因CDKN1A促进颗粒细胞增殖，且外源性胰岛素刺激能够高表达miR-93。为深入探讨miR-93介导胰岛素在PCOS卵巢局部中的作用及其机制，本项目拟进一步筛选胰岛素刺激下调控miR-93的转录因子，并通过EMSA、CHIP、BSP等方法获得转录因子调控miR-93的直接证据，揭示胰岛素在转录和表观遗传学水平调控miR-93表达，进而参与卵巢局部生物学功能的分子机制；为阐明PCOS高胰岛素血症调控卵巢功能内在作用关键因子，阻断其病理进程提供新线索；为临床治疗提供新思路，为发展新型基因治疗药物奠定基础。

卵巢功能障碍、胰岛素抵抗是多囊卵巢综合征（PCOS）重要的病理特征之一，胰岛素参与了卵巢功能的调控，但具体机制不明确。本课题组前期研究证实胰岛素抵抗状态下microRNA调控卵巢功能的关键分子。我们前期筛选和验证了PCOS卵巢局部miR-93高表达，证实miR-93通过其靶基因细胞周期相关基因CDKN1A促进颗粒细胞增殖，且外源性胰岛素刺激能够高表达miR-93。为深入探讨miR-93介导胰岛素在PCOS卵巢局部中的作用及其机制，本项目进一步探讨了PCOS卵巢局部功能障碍的机制，结果显示miR-93、高雄激素能够调控AMH的表达，基因多态位点rs13405728与卵巢反应性异常相关。同时从胰岛素信号通路出发，证实了胰岛素能够调控miR-93的表达，lncRNA与miR-93可能存在一定的调控机制，但具体机制仍不清晰。揭示胰岛素在转录和表观遗传学水平调控miR-93表达，进而参与卵巢局部生物学功能的分子机制；为阐明PCOS高胰岛素血症调控卵巢功能内在作用关键因子，阻断其病理进程提供新线索；为临床治疗提供新思路，为发展新型基因治疗药物奠定基础。