外泌体来源miRNA-221调节小鼠子宫内膜细胞EMT及养血安胎方对RSA作用机制的实验研究

复发性流产(RSA)是常见的生殖障碍性疾病，胚胎植入过程中，子宫内膜发生上皮间充质转化（EMT），RSA的发生与该过程减弱密切相关。前期实验发现我院内部制剂养血安胎方通过干预EMT过程对RSA起到关键治疗作用，并引起miRNA-221为主的多种与EMT相关的外泌体miRNA变化。据此，我们提出假说：子宫内膜上皮和基质细胞分泌的外泌体中可能存在多种miRNA参与EMT，并以miRNA-221为主；通过养血安胎方干预外泌体来源miRNA-221靶向转录因子调控EMT，达到治疗RSA作用。故本项目将利用miRNA芯片技术筛选并验证外泌体来源miRNA对子宫内膜EMT的调控作用；利用RNA-seq和ATAC-seq技术通过动物实验和细胞实验阐明养血安胎方通过外泌体促进EMT对RSA的治疗作用，明确养血安胎方通过miRNA-221相关转录因子调控EMT的作用机制，为养血安胎方治疗RSA提供科学依据。

Recurrent spontaneous abortion (RSA) is a common reproductive disorder. During embryo implantation, the endometrium underwent epithelial mesenchymal transformation (EMT), and the occurrence of RSA is closely related to the weakening of this process. Preliminary experiments found that the internal preparation " Yangxue antai formula" in our hospital played a key therapeutic effect on RSA by interfering in the EMT process, and caused a variety of miRNA changes of EMT-related exosomes, mainly miRNA-221. Based on this, we proposed a hypothesis: exosomes secreted by endometrial epithelial cells and stromal cells may have a variety of miRNA participating in EMT, mainly miRNA-221.The therapeutic effect on RSA was achieved through the intervention of Yangxue antai formula to regulate EMT by targeting transcription factors of miRNA-221 from exosomes.Therefore, this study intends to use miRNA chip technology to screen and verify the regulatory effect of exosome source miRNA on endometrial EMT. RNA-seq and ATAC-seq technologies were used to elucidate the therapeutic effect of Yangxue antai formula on RSA by promoting EMT through exosomes through animal experiments and cell experiments, and to clarify the mechanism of Yangxue antai formula's regulation of EMT through miRNA-221 related transcription factors, so as to provide scientific basis for the treatment of RSA by Yangxue antai formula.