雄激素受体(AR)调控Beclin1致自噬亢进在PCOS卵泡凋亡障碍中的作用及机制研究

Polycystic ovary syndrome (PCOS), characterized by hyperandrogenism and abnormal follicular development, is the most leading cause of anovulatory infertility that severely threatens women’s reproductive and metabolic health. Granulosa cells (GCs) are believed to play key roles in follicular development, maturation or atresia. Our previous published study showed that GCs from PCOS women presented apoptosis disorder. Furthermore, DHT-treated GCs (in vitro) and GCs from testosterone propionate induced PCOS rats (in vivo) also showed significant increase in apoptosis. However, the mechanism remains to be illustrated. Our group found that GCs from PCOS women had increased number of autophagosome, with up-regulated expression of Beclin1, a central regulator of autophagy. Based on the above outcomes, we will investigate the transcriptional mechanism of which androgen/AR promote the transcription of the Beclin1 by luciferase assay and chromatin immunoprecipitation assay. With RNAi and adenovirus-mediated overexpression, we will evaluate the effects of Beclin1 expression on apoptosis dysfunction and related molecular mechanisms. The current project is intended to discuss the regulating role of AR mediated autophagy dysfunction in follicular apoptosis disorder and provide theory basis for clinical intervention of PCOS.

以高雄和卵泡发育障碍为特征的多囊卵巢综合征(PCOS)是不排卵性不孕的最常见病因，严重影响女性健康。颗粒细胞(GC)在调控卵母细胞发育、成熟或凋亡中发挥至关重要的作用。课题组前期已发表论文揭示PCOS患者卵泡颗粒细胞凋亡障碍，且体外细胞实验及PCOS大鼠模型也证实高雄暴露可引起卵泡颗粒细胞凋亡障碍，但其机制尚未明确。进一步研究发现以高雄症为主要病理生理的PCOS颗粒细胞中自噬小体数量显著增加，伴随着自噬启动，相关执行基因Beclin1亦显著上调。基于此，本课题拟以PCOS颗粒细胞中Beclin1为切入点，利用临床标本和体内外实验，采用荧光素酶报告系统、染色质免疫共沉淀、RNAi及腺病毒过表达技术明确雄激素通过雄激素受体对Beclin1的转录调控作用，并阐明Beclin1的异常高表达在PCOS卵泡发育障碍中的重要作用及其分子机制。研究结果为PCOS的病理生理机制和临床诊治奠定坚实的理论基础。

以高雄和卵泡发育障碍为特征的多囊卵巢综合征(PCOS)是排卵障碍性不孕的最常见病因，严重影响女性的生育及健康。颗粒细胞(GC)在调控卵母细胞发育、成熟或凋亡中发挥至关重要的作用。课题组前期已发表论文揭示PCOS患者卵泡颗粒细胞凋亡障碍，且体外细胞实验及PCOS大鼠模型也证实高雄暴露可引起卵泡颗粒细胞凋亡障碍，但其机制尚未明确。本课题利用临床样本证实以高雄症为主要病理生理的PCOS颗粒细胞中自噬小体数量显著增加，伴随着自噬启动，相关执行基因Beclin1亦显著上调，证实PCOS卵巢颗粒细胞存在自噬亢进；体外细胞实验证实高雄在这一过程发挥重要作用。本研究结果为PCOS的病理生理机制和临床诊治奠定坚实的理论基础。.此外，有研究提示正五聚蛋白3（PTX3）在多囊卵巢综合征发生中发挥重要作用，但是目前的结果有待争议。本项目开展收集PCOS女性卵泡液，证实PCOS女性卵巢局部微环境中PTX3显著升高，且在外周血中亦证实该现象，提示PCOS外周循环及卵巢局部存在固有免疫水平异常，进一步相关性分析发现PTX3且与高雄症的病理生理密切相关，该研究从固有免疫异常角度开辟PCOS高雄症发生发展的新机制。