高雄激素环境通过AR-PI3K/Akt信号通路对PCOS患者子宫内膜FOXO1的调控及影响胚胎种植的机制研究

Polycystic ovary syndrome (PCOS) is a common disease of reproductive age with features of hyperandrogenism. Our previous studies and recent literatures showed that, in addition to ovulation disorders, abnormal functions of endometrium may also be an important reason for the decline in pregnancy rate and the increased miscarriage rate in patients with PCOS. However, the mechanism by which androg endometrial function and embryo implantation in patients with PCOS, so as to improve the endometrial receptivity, and finally to increase the embryo implantation rate and to reduce the early miscarriage rate. FOXO1 (Forkhead box class O-1) is an important transcription factor in the regulation of apoptosis, differentiation, glucose metabolism and oxidative stress. Our recent research with bioinfomatic analyses and pre-experiments found that the local FOXO1 expression in endometrium of patients with PCOS was decreased, suggesting that hyperandrogenic environment may influence the endometrial receptivity during the process of embryo implantation in patients with PCOS through AR-PI3K/Akt-FOXO1 pathway. The present research project will use methods combined with in-vivo and in-vitro experiments of human endometrial cells, following with the establishment of 3D implantation model by co culturing human endometrium tissue with human blastocysts diagnosed with severe thalassemia, so as to explore the mechanisum of hyperandrogenism environment in control of the endometrial function and implantation effect through FOXO1 and its downstream markers. The results of our research project will provide experimental basis and new markers of the anti-androgenic therapy in patients with PCOS, in order to finally improve their clinical pregnancy outcomes.

多囊卵巢综合征是以高雄激素为主要特征的育龄期常见病。我们的前期研究及近期文献报道PCOS患者的子宫内膜异常亦是其妊娠率下降及流产率增加的重要因素。雄激素影响子宫内膜功能及胚胎种植的机制尚未明确；深入探讨其机理对提高PCOS患者的内膜容受性，改善妊娠结局有重要意义。FOXO1是子宫内膜中调节分化、凋亡、糖代谢及氧化应激等重要功能的转录因子。我们的预实验提示，高雄激素的PCOS患者内膜局部FOXO1表达下降，结合生物信息学预测结果，高雄激素环境可能通过AR-PI3K/Akt信号通路，在转录和翻译后修饰水平调控内膜间质细胞FOXO1的表达和功能，导致其稳态失衡，进而影响胚胎种植。本课题拟采用人子宫内膜进行体内外细胞实验，并利用种植潜能正常的人类重型地贫囊胚构建体外三维种植模型，阐明高雄激素环境对PCOS患者子宫内膜FOXO1的调控机制及胚胎种植效应，为临床抗雄激素治疗提供新的实验依据及检测标志物

近期研究发现，PCOS患者除排卵异常外，其子宫内膜异常亦可能是其妊娠率下降及流产率增加的原因之一。探讨PCOS患者子宫内膜异常的发生机制及明确其对胚胎种植的影响，可能是提高PCOS患者胚胎种植率和继续妊娠率的关键环节，对改善其妊娠结局可能具有重要意义。.FOXO1是调节增殖和凋亡、分化、糖代谢、氧化应激等功能的重要转录因子，并主要受PI3K/Akt信号通路的调控。目前研究发现，FOXO1主要在子宫内膜间质中表达，调节子宫内膜局部代谢、凋亡、分化、蜕膜化等，并可能受胰岛素信号调控。高胰岛素血症是PCOS患者常见的内分泌代谢紊乱特征之一。PCOS患者的高胰岛素环境是否通过调控FOXO1的表达和功能导致子宫内膜局部的代谢、分化、蜕膜化及容受性等方面的异常？另外，PCOS患者特征性的高雄激素环境对子宫内膜局部FOXO1是否具有调控作用？以上问题的答案目前尚无研究证实。.本研究通过临床结合基础研究，旨在探讨以下几个科学问题：1. 在最大限度地匹配和平衡了卵子和胚胎因素后，PCOS患者进行辅助生殖技术后的临床妊娠率、早期妊娠丢失率及活产率与非PCOS患者相比是否有差异？子宫内膜功能在多大程度上参与了PCOS患者不良妊娠结局的发生？2. 高胰岛素环境及高雄激素环境是通过何种机制影响和调控PCOS患者子宫内膜间质细胞FOXO1的表达及功能，并最终导致其下游包括蜕膜化、增殖分化、容受性等多方面功能的失调？.本课题共分为三个部分：1、回顾性配对病例对照研究：对进行PGD的PCOS和非PCOS患者进行1:3匹配年龄、BMI、胚胎评分，比较两组患者单胚胎移植后的临床妊娠结局。2、为生物信息学分析，旨在筛选胰岛素/雄激素对子宫内膜FOXO1的调控通路及下游效应分子网络。3、机制研究，在第二部分结果的基础上，通过子宫内膜细胞实验，探讨高胰岛素环境/高雄激素环境通过PI3K/Akt信号通路对子宫内膜间质细胞FOXO1的调控机制及对其下游包括蜕膜化、增殖分化、容受性等标志物表达的影响，及PCOS患者子宫内膜间质细胞中PI3K/ Akt/ FOXO1通路的改变和效应。