精子组蛋白异常保留区印记基因PEG3在原因不明复发性流产中的作用机理

精子表观遗传信息对于早期胚胎的发育有着重要的作用，组蛋白保留区印记基因的甲基化错误可以传递到子代影响胚胎的发育并导致早期流产的发生。本课题前期实验发现原因不明复发性流产患者配偶的精子组-精蛋白转换异常，印记基因PEG3启动子甲基化异常，携带异常表观遗传信息的精子可能导致胚胎发育异常。本课题拟探究精子组蛋白异常保留区的差异印记基因，进一步验证PEG3的甲基化异常影响子代胚胎绒毛发育，异常的绒毛将导致流产的发生机制。主要通过染色质免疫共沉淀联合启动子芯片技术来筛查精子组蛋白异常保留区的差异印记基因，焦磷酸测序检测精子和流产绒毛组织印记基因PEG3甲基化水平。在动物水平，探索异常的印记基因对胚胎发育的影响，通过过表达/沉默印记基因PEG3探究其具体的作用机制。本研究能丰富复发性流产的病因学，重新评估男性精子表观遗传因素在复发性流产中的作用，为该疾病的诊断和治疗提供一定的理论依据。

The epigenetic information of sperm plays an important role in the early development of embryos.Histone retained at imprinted genes and their DNA methylation errors could be passed to offspring in affected embryonic development and led to early miscarriage.In our pre-experiments,we found that the sperm histone-protamine alternation and methylation level of imprinted gene PEG3 were abnormal in unexplained recurrent spontaneous abortion (RSA) group.The sperm carried the abnormal epigenetic information might lead to abnormal embryonic development and cause abortion.The research intends to explore that sperm imprinted genes of abnormal histone retained and the methylation levels of PEG3 will transmitted to the progeny embryos,how dysplasia fluff will lead to the occurrence of miscarriage. We will screen imprinted genes of abnormal histone retained through chip combined with promoter microarray analysis. Pyrosequencing detected the methylation levels of imprinted genes in sperm and abortion villi.In animal level,we will discuss aberrant imprinted genes on the development of embryos.We will study the mechanism by overexpression or silencing of imprinted gene PEG3.The research can enrich the etiology and re-evaluate the role of sperm epigenetic factors in RSA. Aslo,it can provide some theoretical basis for the diagnosis and treatment of this disease.