GnRH类似物通过垂体外作用影响子宫内膜容受性的机制研究

Gonadotropin releasing hormone analogues (GnRHa) have been widely utilized in assisted reproduction techniques（ART）, wheras the mechanism of their effect in lowering endometrial receptivity is poorly understood. Our research group preliminary study showed that GnRH receptor (GnRHR) expresses in endometrium, and its level differs in implantation window stages, suggesting that GnRH/GnRHR system directly functions in endometrium via extrapituitary. Therefore, we propose the hypothesis that via extrapituitary GnRHa directly regulate GnRH/GnRHR system, resulting to endometrial receptivity defects. In order to testify this, we intend to do the research in the following aspects: ① Collecting patient samples (both proliferative phase and secretory phase), and detect GnRH, GnRHR and its subtypes, intergrin av3, FAK, PyK2 and its phosphorylation type etc expression level. ② By comparing the GnRH analogue treatment group with solvent treatment group in 3D model by protein chip, find out the possible downstream singnal pathway, and using western blot to validate these results. ③ By pituitary detrimental mice, further validate the molecular mechanism in animals. This research would provide new insights into extrapituitary mechanism of GnRH/GnRHR in uterine endothelium, and theoretical basis in optimizing ART for clinical usage.

促性腺激素释放激素类似物（GnRHa）在辅助生殖中广泛应用，但其影响子宫内膜容受性的深入机制尚不清楚。课题组前期研究结果显示人子宫内膜表达GnRH受体(GnRHR)，且在种植窗不同阶段有表达差异。基于此本课题提出如下假设：GnRHa通过垂体外机制直接作用于子宫内膜中GnRHR/整合素信号通路，影响子宫内膜容受性。本课题拟行以下研究：①检测不同时期（增生期、分泌期）内膜标本GnRHR、FAK和PyK2表达，及内膜容受性标志物整合素等表达水平；②应用GnRHa干预3D内膜共培养实验系统，蛋白质芯片对比找出GnRH可能的下游信号通路，尤其是GnRHR/FAK/PyK2/integrin-av3通路的作用；③利用垂体受损小鼠进一步证实GnRH的下游通路分子机制。本研究结果将对阐明GnRH/GnRHR在子宫内膜容受性中的垂体外作用机制，为提升胚胎种植率提供理论基础。

促性腺激素释放激素（GnRH）类似物促排卵等作用在辅助生殖中已广泛应用。但是GnRH类似物影响子宫内膜容受性，其深入机制尚不清楚。研究结果显示人子宫内膜表达GnRH受体(GnRHR)，且在种植窗不同阶段具有表达差异。GnRH类似物是否通过垂体外机制直接作用于子宫内膜组织中的GnRH/GnRHR信号通路，影响子宫内膜容受性？本课题研究：提取了子宫内膜原代上皮细胞和基质细胞，基本构建了子宫内膜3D模型。通过体外模拟胚胎着床的粘附实验，GnRH类似物预处理的内膜细胞PCR与Western blot结果表明GnRH类似物不显著影响子宫内膜细胞GnRHR/integrin avβ3-FAK/PyK2信号通路，蛋白组学检测亦表明GnRH类似物直接作用于子宫内膜细胞时，对主要的内膜容受性标志分子不产生显著影响。