PATL2调控ADM2抑制颗粒细胞凋亡在PCOS卵泡异常发育中的作用及机制研究

已知颗粒细胞凋亡增加是导致PCOS卵泡异常发育的重要因素，但相关调控机制尚不完全清楚。申请者预实验表明：PCOS患者卵巢颗粒细胞mRNA绑定蛋白PATL2表达减少与颗粒细胞凋亡密切相关。结合生物信息学预测，我们假设PATL2可能通过稳定ADM2的mRNA水平，增加ADM2表达，进而发挥抑制颗粒细胞凋亡和促进卵泡正常发育的作用。因此，本项目拟利用两种人卵巢颗粒细胞系，结合RT-PCR，免疫印迹，RIP，质粒转染和Crispr/Cas9敲除基因等技术方法，围绕PATL2增加ADM2表达及其抑制颗粒细胞凋亡的分子机制开展研究；随后，利用卵巢特异性敲除PATL2基因小鼠建立PCOS动物模型，进一步证明PATL2在PCOS卵泡异常发育中的作用与机制，为深入理解PCOS患者卵泡异常发育的病理机制提供新的理论依据，并期望为改善临床PCOS患者的卵泡质量开辟新的诊疗方法，具有重要理论意义和潜在应用价值。

It has been reported that increased level of granulosa cells apoptosis is an important factor to result in the formation of PCOS related abnormal follicles. Based on the previous findings, we hypothesis that in PCOS ovaries, reduced expression of PATL2 in granulosa cells might promote granulosa cell apoptosis by down-regulating ADM2 expression, which disturbing the normal follicular development and leading to the formation of PCOS related aberrant follicles. Thus, we plan to explore the regulatory mechanism of PATL2 regulating ADM2 expression to affect granulosa cells apoptosis by using two different human ovary granulosa cell lines and RT-PCR, Western blot, RIP, gene over expression and Crispr/Cas9 knockout, et al. furthermore, we plan to further confirm the in vitro outcomes by using PATL2 gene ovarian specificity knockout mice to establish the PCOS mouse model. This study will help us to better understand the regulatory mechanism of granulosa cell apoptosis in PCOS aberrant follicles, and provide valuable guidance and experiment data for increasing the quality of PCOS related follicles and benefiting IVF clinical outcomes.