外泌体在母胎交叉对话中的潜在作用研究

Embryo implantation is a complicated process. Not only the embryo with implantation competency but also the receptive endometrial are required. Meanwhile, they need to developed at the a synchronized stage. It is a complex and interesting event during the process of synchrony as development of the embryo and the endometrium are two separated procedures. The maternal and the embryo may communicate with each other through molecular factors to facilitated the embryo adhesion and implantation. Exosomes are nanometer sized extracellular vesicles. The contents of exosomes including DNA, RNA, proteins and lipids. These cargos could be transferred to other cells and exert functions inside the target cells. At this moment the function of exosomes in maternal and embryo cross talk are not well studied. It is hypotheses that the endometrial cell could secret the exosome which contained miRNAs. Those exosomes could transfer the miRNAs to the embryo and regulated its development and implantation. This project will study the regulation function of estrogen and progesterone on the miRNA expression in endometrial cell derived exosomes. Moreover, the mediation function of exosome in maternal and embryo cross talk will be investigated. Lastly, the prediction roles of exosomes on endometrial receptivity will also be explored. The results of this project could provide more information about the crosstalk between the embryo and the mother and help us to understand the regulatory role of the uterus to the embryo which may increase the successful rate of in vitro fertilization(IVF).

胚胎着床是一个复杂的过程，需要具有发育潜能的优质胚胎和处于接受态的子宫内膜的同步发育并协调互动，胚胎与母体通过细胞因子交流对话，协调互作完成粘附和植入等一系列重要步骤。外泌体为细胞分泌的纳米级囊泡，其所携带的核酸、蛋白等分子赋予外泌体丰富的生物学信息，其能够携带RNA(如干扰RNA、微小RNA)，通过细胞膜融合将其内含物传送入靶细胞的内部，在转录水平及转录后水平调控靶细胞的生物学行为。目前外泌体在母胎交叉对话中的作用尚未完全阐明，推测子宫内膜细胞可分泌携带miRNA的外泌体，并可将miRNA等生物信息传递到胚胎中，从而影响胚胎的发育及着床。子宫内膜外泌体还可能反应子宫内膜的容受性状态。本项目将通过研究雌孕激素对子宫内膜外泌体的调节作用，检测其携带miRNA的变化以及功能，探讨外泌体在母胎交叉对话的作用，并了解其对子宫内膜容受性的预测，为其临床应用做前期基础研究，以提高辅助生殖成功率。

目的：探究外泌体在母胎交叉对话中的作用机制。方法：1.通过超速离心的方法分离人宫腔液中的外泌体及子宫内膜细胞系ishikawa细胞的外泌体。2.通过外泌体-细胞共培养模型、胚胎体外黏附模型及小鼠模型等实验，验证子宫内膜外泌体对胚胎的作用机制。3.通过新一代测序技术，比较接受辅助生殖助孕的胚胎反复着床失败患者与胚胎可正常着床女性胚胎着床期宫腔液中外泌体miRNA的表达。结果：1.可通过超速离心的方法分离出人宫腔液中外泌体，此类外泌体可表达CD63及HSP70，人宫腔液中外泌体的直径平均为223.2±5.3nm。2.子宫内膜来源的外泌体可进入胚胎细胞中，并可将miRNA携带进入胚胎细胞并抑制胚胎细胞中靶基因的表达。3.通过抑制子宫内膜细胞产生的外泌体，可以降低胚胎的着床率。4.反复着床失败患者的子宫腔液中外泌体miRNA的表达与对照组存在显著差异，生物信息学分析显示其可能通过细胞黏附等通路影响胚胎着床。结论：母胎交叉对话及是一个复杂而动态的过程。子宫内膜外泌体可通过miRNA参与到母胎交叉对话中并影响胚胎着床。本项目对于子宫内膜外泌体的研究可以扩展我们目前对生殖生理学和病理学的理解和认识。子宫内膜外泌体，特别是子宫内膜外泌体中的miRNA可以作为干预目标或预测疾病和辅助生殖助孕结果的生物标志物，期待更多关于子宫腔液外泌体的研究以进一步验证及推广。