抗苗勒氏管激素通过cAMP/CREB调控子宫内膜蜕膜化的分子机制

Decidualization is characterized as an important feature of implantation and essential for pregnancy. The marker of endometrial decidualization, insulin-like growth factor binding protein-1 (IGFBP1),is effectively regulated by cyclic adenosine monophosphate(cAMP) pathway. To our knowledge the current study reports AMH is able to inhibit the differentiation and proliferation of endometrial stromal cells (ESC) in decidualization.We have found that AMH canonically propagates its actions via cAMP signaling in granulose cells. Our preliminary study suggested that AMH down-regulates IGFBP1 in vitro by using gene cloning , therefore, we hypothsize that AMH plays an important role in decidualization. To test this hypothesis, we will investigate the mechanisms that how AMH inhibits the expression of IGFBP1 through the cAMP/CREB signal pathway by using Chromatin Immunoprecipitation,EMAS and Luciferase Report Gene. Additionally, we will empoly a novel mouse Endometrial Three-Dimensional (3D) Culture System to mimic the human endometrium, and demonstrate the failure of embryo adhesion, invasion and implantation due to abnormal decidualization. Meanwhile, to indicate the effect of AMH on decidualization in vivo,we will induce decidualization of mouse endometrium and transfer the embryo to the mouse uterine, then observe the rate of implantation, miscarriage and live birth. This study will reveal a potential regulatory mechanism in decidualization which may develop new therapies for reproductive diseases such as infertility，abortion and recurrent implantation failure.

子宫内膜蜕膜化是正常着床和妊娠的一个重要特征,cAMP通路是调控蜕膜化标志物IGFBP1的重要途径。文献报道抗苗勒氏管激素(AMH)通过抑制子宫内膜间质细胞(ESC)的增殖分化参与蜕膜化过程,但机制不清。我们在颗粒细胞中研究证实AMH通过cAMP途径发挥生物学效应；前期工作发现:AMH负调控ESC细胞中IGFBP1的表达，因此推测AMH可能通过cAMP通路调控子宫内膜蜕膜化。本项目拟采用免疫共沉淀、EMSA、荧光素酶报告基因阐明AMH通过cAMP/CREB途径调控IGFBP1的分子机制；利用体外三维类子宫内膜-鼠胚共培养体系,进一步证实AMH通过调控内膜蜕膜化进而影响胚胎黏附、侵入及着床；通过小鼠在体移植术，研究靶向干扰子宫内膜AMH对鼠胚着床率、流产率及活产率的影响；探讨AMH调控IGFBP1分子机制在子宫内膜蜕膜化中的意义,为因蜕膜化异常引起的不孕、反复种植失败、流产提供新的诊治思路。

RIF是导致女性不孕的常见内分泌疾病，病因不明。母胎界面关键蛋白和激素的异常表达影响母胎沟通，导致不良妊娠结局。观察抗苗勒氏激素(AMH)及其Ⅱ型受体(AMHRII)在子宫内膜的表达。本研究旨在研究AMH和AMHRII在人子宫内膜、蜕膜组织和囊胚中的表达。采用免疫组织化学和实时荧光定量PCR检测AMH和AMHRII在RIF患者中的表达。结果表明，AMH和AMHRII在正常子宫内膜均有表达，AMHRII在黄体中期高表达。此外，AMHRII在整个妊娠过程中均有表达，且在妊娠中期表达量最高。AMHRII在囊胚中有表达，但未观察到AMH。AMHRII在RIF组子宫内膜中的阳性表达率明显高于RIF组。在AMHRII高表达的RIF中，雌激素受体(ER)、胰岛素样生长因子结合蛋白1(IGFBP1)和催乳素(PRL)的表达明显降低。AMHRII高表达患者的细胞凋亡率明显高于AMHRII正常表达患者。我们的数据提示AMHRII通过AMH和AMHRII信号通路对RIF产生影响。它通过干扰子宫内膜蜕膜化和细胞凋亡参与RIF的发生发展。探讨AMH调控IGFBP1分子机制在子宫内膜蜕膜化中的意义,为因蜕膜化异常引起的不孕、反复种植失败、流产提供新的诊治思路。