N-糖基化修饰的骨桥蛋白（OPN）参与胚胎发育与着床调控及其作用机制研究

Osteopontin(OPN) is N-connection glycoprotein,one of most important cell extracellular matrix (ECM) components ,which resides in many tissues and organs. It also regulates cell adhesion, migration, differentiation and so on. However, reportsofmechanisms involved in embryonic development and recognition of uterus-embryoarelimited. The immunofluorescence, Western blot, and Real-time PCR technology are applied in the study.①Wewant to compare the OPN expression in differential cleavage stage of mouse embryosand uterine endometrium in peri-implantation.②The location and expression of OPN are analyzed in the endometrium of proliferative phaseand the secretory phase.③The factor andhormone, which affect the expression of OPN, embryonicadhesion, and implantation,was discovered by using the model of in vivo and in vitromouse implantation. ④We transferredOPN plasmids into endometrial cells and analyzedthe influence of embryo implantation and development that wereinduced by OPN.⑤Analysis of regulatory function of OPN mediated cell identification and molecular action mechanisms of adhesion and signaling pathways.⑥Usinglectins Mal andConA,in addition to methods such as mass spectrum,to analyze the function of the function of OPN in using sugar as a target to interfere with pregnancy in clinical practice.OPNregulates the expression of embryo implantation related factors. This is observed by the gene interference, the molecular mechanism of recognition, and the adhesion of uterus-embryo mediated by OPN regulating embryonic development is revealed, through several pathways.

骨桥蛋白（OPN）为N连接糖蛋白，作为细胞外基质成分之一，存在于多种组织器官，介导细胞黏附、迁移、分化等，但参与胚胎发育、调控母-胎间识别机制研究极少。本研究拟用免疫荧光、Western blot、Real-time PCR等技术①分析不同卵裂阶段小鼠胚胎及围着床期子宫内膜OPN表达规律；②分析临床增生期、分泌期子宫内膜OPN表达与定位；③借助体内（外）着床模型对比分析激素及着床相关因子对OPN表达影响及对胚胎与子宫内膜识别黏附能力的影响；④构建OPN表达调控质粒转染模拟高低接受态的子宫内膜细胞，分析调控OPN表达对胚胎发育及着床能力的影响；⑤ 分析OPN参与胚胎发育与着床调控作用机制及信号转导通路；⑥以凝集素MAL、ConA以及质谱等方法分析OPN功能的糖链修饰作用。揭示N连接糖蛋白OPN调控胚胎发育、介导胚胎与子宫内膜识别、黏附的分子机制，为临床避孕和助孕干预提供新思路。

骨桥蛋白（OPN）为N连接糖蛋白，作为细胞外基质成分之一，存在于多种组织器官，介导细胞黏附、迁移、分化等，但参与胚胎发育、调控母-胎间识别机制研究极少。本研究拟用免疫荧光、Western blot、Real-time PCR等技术①分析不同卵裂阶段小鼠胚胎及围着床期子宫内膜OPN表达规律；②分析临床增生期、分泌期子宫内膜OPN表达与定位；③借助体内（外）着床模型对比分析激素及着床相关因子对OPN表达影响及对胚胎与子宫内膜识别黏附能力的影响；④构建OPN表达调控质粒转染模拟高低接受态的子宫内膜细胞，分析调控OPN表达对胚胎发育及着床能力的影响；⑤ 分析OPN参与胚胎发育与着床调控作用机制及信号转导通路；⑥以凝集素MAL、ConA以及质谱等方法分析OPN功能的糖链修饰作用。揭示N连接糖蛋白OPN调控胚胎发育、介导胚胎与子宫内膜识别、黏附的分子机制，为临床避孕和助孕干预提供新思路。围绕相关研究，我们发表了17篇SCI论文，参加了学术交流并发表了4份报告，培训了25名研究生，并获得了2项科学技术奖。