GRIM-19在胚胎着床中的作用及其机制研究

Embryo implantation is the key to successful pregnancy, and its molecular mechanism is the focus of study in the field of reproductive medicine. As cellular energy metabolism and apoptosis regulation centers, the mitochondrial play important roles in the process of embryo development and implantation. GRIM-19 is a newly discovered cell apoptotic regulation genes. As a functional component of mitochondrial complex I, GRIM-19 is required for the assembly of the complex and the mitochondrial electron transfer chain. Our previous study demonstrated that the expression of GRIM-19 is high regulated in the window of uterine receptivity. Further experiments showed that the implantation rate of GRIM-19+/- mouse was significantly lower than that of GRIM-19 +/+ mouse. The results mentioned above suggest that GRIM-19 play an important role in the implantation process. In vivo and in vitro study, GRIM-19+/+ and GRIM-19 +/- mice were used to explore the effect and mechanism of GRIM-19 on implantation by evaluating mitochondrial structure and function, cell proliferation and apoptosis, endometrial receptivity, angiogenesis and the interactivity of STAT-3 and HIF-1α. Then, we are intent to study the relationship between the expression of GRIM-19 and human endometrial receptivity, and to explore the role of GRIM-19 in implantation failure. The findings of these studies may provide important theoretical values for exploring the molecules mechanism of mitochondrial protein in implantation, and useful experimental foundations for early prediction and targeted therapy of implantation failure.

线粒体在胚胎着床中发挥重要作用，GRIM-19是参与调控细胞凋亡和线粒体呼吸作用的基因，作为线粒体呼吸链复合物I的一部分，对复合体的装配和线粒体电子转移链是必需的。本项目组的前期研究发现，GRIM-19在小鼠着床的"窗口期"表达增强，且GRIM-19+/-小鼠胚胎着床率明显降低，因此推测GRIM-19在着床过程中发挥重要作用。本课题以GRIM-19+/+和GRIM-19+/-小鼠为研究对象，建立体内和体外着床模型，研究GRIM-19对线粒体、细胞凋亡、子宫内膜容受性和血管新生的影响，并深入研究GRIM-19在着床中对STAT3和HIF-1α的调控作用和机制。在此基础上，研究GRIM-19与人子宫内膜容受性的关系，并进一步探讨GRIM-19在胚胎着床失败中的作用。本课题的研究对于阐明线粒体蛋白在着床中的分子作用机制具有重要的理论价值，并为着床失败的早期预测和靶向治疗提供新的思路和实验依据。

课题组按照研究计划如期完成了本课题的研究工作。本课题以GRIM-19+/+和GRIM-19+/-小鼠为研究对象，建立体内和体外着床模型，研究GRIM-19在小鼠着床窗口期对线粒体功能、细胞凋亡、子宫内膜容受性等的调控。在此基础上，研究GRIM-19与人子宫内膜容受性的关系。项目取得成果：（1）GRIM-19在小鼠早期妊娠第1-6天子宫腔上皮和腺上皮均有表达，妊娠第4天小鼠子宫内膜GRIM-19蛋白及mRNA表达水平降低，且凋亡减少；（2）体内着床模型中GRIM-19+/-小鼠胚泡着床率明显低于GRIM-19+/+小鼠；（3）体外着床模型中，过表达GRIM-19后，RL95-2-BeWo 共培养球状体黏附率降低，线粒体膜电位降低，细胞凋亡增加；（4）过表达GRIM-19后，RL95-2细胞系中STAT3及IL-11 蛋白和mRNA水平降低，TNF-α蛋白和mRNA水平升高。研究结果表明GRIM-19通过调节细胞凋亡、胚胎黏附和免疫耐受在早期胚胎着床中发挥作用。本课题的研究对于阐明线粒体蛋白在着床中的分子作用机制具有重要的理论价值，并为着床失败的早期预测和靶向治疗提供新的思路和实验依据。在完成计划研究内容的基础上，课题组对项目研究进行了拓展延伸，研究了GRIM-19在稽留流产及子宫腺肌病中的作用及相关机制。研究证实GRIM-19在早期妊娠中起重要作用，GRIM-19的低表达可能通过线粒体介导的细胞凋亡和减少新生血管生成导的稽留流产的发生。研究还证实GRIM-19可通过GRIM-19-STAT3-VEGF轴调节血管生成和细胞凋亡促进了子宫腺肌病的发生。