富亮氨酸糖蛋白-1(LRG-1)激活TGF-β/Smad通路促进植入前胚胎发育的机制研究

The potential of preimplantation embryo development is one of the important factors affecting the success rate of in vitro fertilization - embryo transfer (IVF - ET), and is also a main reason for deciding healthy offspring. Through proteomic high-throughput screening, we found that the content of Leucine-rich-alpha-2-glycoprotein-1 (LRG-1) in the culture medium of embryos in the implantation group was significantly higher that in the non- implantation group, prompting that LRG - 1 may be one of the specific molecules of high quality embryos. In this study, it was proposed that the Time-lapse imaging, siRNA interference, plasmid transfection and other techniques be used to further explore important regulation function of LRG - 1 for the potential of preimplantation embryo development, and explore the molecular mechanism of promoting embryonic development by LRG - 1 activating TGF - beta/Smad pathway, in order to establish a Time - lapse imaging embryo joint assessment system with LRG - 1 as auxiliary index, thus to seek for new ideas and new targets for the application of assisted reproductive technologies.

植入前胚胎发育潜能是影响体外受精-胚胎移植成功率的重要因素，也是决定后代健康的主要原因。我们通过蛋白质组学高通量筛选发现，种植组胚胎的培养液中富亮氨酸糖蛋白-1(LRG-1)含量显著高于未种植组，提示LRG-1可能是优质胚胎的特异性分子之一。本研究拟运用Time-lapse imaging、siRNA干扰及质粒转染等技术进一步探讨LRG-1对植入前胚胎发育潜能的重要调控作用，并探索LRG-1激活TGF-β/Smad通路促进胚胎发育的分子机制，以期建立以LRG-1作为辅助指标的Time-lapse imaging胚胎联合评估体系，为辅助生殖技术的应用寻找新思路和新靶点。

植入前胚胎发育潜能是影响体外受精-胚胎移植成功率的重要因素，也是决定后代健康的主要原因。我们前期研究发现:应用蛋白质组学高通量筛选，富亮氨酸糖蛋白-1(Leucine-rich-alpha-2-glycoprotein-1，LRG-1)为种植组与非种植组的差异蛋白之一，并通过ELISA得以验证。本研究运用外源性添加、免疫荧光染色技术、敲低实验及敲除小鼠生育实验方法，得出以下结论：1）人类优质胚胎与非优质胚胎中，LRG-1表达有显著性差异；2）外源性添加LRG-1不能降低小鼠胚胎的2细胞阻滞，对囊胚形成无显著影响；3）敲除小鼠LRG-1基因，囊胚发育率有显著性下降；4）通过LRG-1基因工程敲除小鼠模型明确LRG-1功能，生育实验显示后代产仔数无显著性差异。综上，在人类胚胎发育过程中，LRG-1可能促进植入前胚胎发育潜能的获得，提高胚胎的种植能力。同时LRG-1可能作为优质胚胎筛选的候选分子之一，联合Time-lapse技术建立胚胎联合评估体系用于优质胚胎的挑选。本课题对提高单胚胎移植的临床妊娠率有重要的参考价值。