复发性流产患者子宫内膜RAGE-NF-κB通路调控EMT改变及养血安胎方作用机制研究

Recurrent abortion (RSA) is a frequently-occurring and difficult disease in women of childbearing age, and the pathogenesis of 30% of patients is still unclear. Regardless of the cause, EMT occurs in the endometrium during embryo implantation, and this abnormality is closely related to the occurrence of RSA. The well-known hospital preparation "Yangxue Antai Decoction" developed by our hospital has a significant effect on the treatment of renal deficiency and blood stasis type RSA. In the early stage, we found that the serum RAGE was down-regulated in patients with RSA through proteomics and other studies. The Yangxue Antai Decoction can up-regulate the expression of RAGE in cells and activate the NF-κB pathway. Based on this, we hypothesized that in the process of embryo implantation in patients with RSA, the RAGE-NF-κB pathway in the endometrial epithelial cells attenuates the EMT process leading to repeated abortion and the Yangxue Antai Decoction can regulate the EMT process to treat RSA. By observing tissue specimens from patients with RSA, this project intends to discover the relationship between EMT attenuating process by the RAGE-NF-κB pathway in the endometrial epithelial cells and repeated abortion; and to demonstrate that the Yangxue Antai Decoction can treat RSA by influencing EMT process through RAGE-NF-κB pathway and elucidate its mechanism by animal experiments, small animal in vivo imaging techniques and cell experiments. This project also intends to provide scientific basis for the method of nourishing kidney, promoting blood circulation and nourishing blood on the treatment of RSA.

复发性流产（RSA）是育龄女性的多发病及疑难病，30%患者发病机制尚不明确。不论何种病因，胚胎植入过程中，子宫内膜发生EMT，该过程异常与RSA的发生密切相关。我院著名院内制剂“养血安胎方”治疗肾虚血瘀型RSA效果显著。我们前期通过蛋白组学等研究发现RSA患者血清RAGE低表达，养血安胎方能够上调细胞内RAGE表达，激活NF-κB通路。据此，我们提出假设：复发性流产患者胚胎植入过程中，子宫内膜上皮细胞RAGE-NF-κB通路减弱EMT过程导致反复流产；养血安胎方可通过该通路调控EMT过程治疗RSA。本项目拟通过观察RSA患者组织标本研究子宫内膜上皮细胞RAGE-NF-κB通路减弱EMT过程与反复流产的关系；并应用动物实验、小动物活体成像技术及细胞实验证明养血安胎方可通过RAGE-NF-κB通路影响EMT过程以达到治疗RSA的效果并阐释其作用机制。本项目将为养血安胎方治疗RSA提供科学依据。

复发性流产（RSA）是育龄女性的多发病及疑难病，多数患者发病机制尚不明确，严重影响患者身心健康。我们前期通过蛋白组学等研究发现RSA患者血清RAGE低表达，养血安胎方能够上调细胞内RAGE表达，激活NF-κB通路。肾虚血瘀证复发性流产患者与正常女性之间，118种细胞因子具有明显差异表达。进一步通过丰度聚类分析，ELISA技术验证筛选出RAGE基因作为本项目研究重点。因此，本课题以子宫内膜细胞EMT和NF-kB信号通路为研究重点，以养血安胎方为研究对象，利用小鼠动物模型，原代细胞培养，蛋白印迹，分子生物学等技术方法探寻养血安胎方可通过RAGE基因和NF-kB通路调控子宫内膜细胞EMT过程治疗RSA。研究结果发现子宫内膜细胞RAGE被激活可引起一系列细胞内源信号通路变化。RAGE与AGEs结合能够激活细胞内NF-κB信号通路。NF-κB是RAGE的主要作用靶点，其活化可以促进一系列炎症细胞因子的表达，这些炎症因子又可反馈性促进NF-κB的的活化。RAGE介导的NF-κB活化是一个正反馈环路，这一特性可以正反馈促进RAGE的表达，使得RAGE下游信号通路的激活得以维持。RAGE表达水平与EMT呈正相关，EMT进一步调控小鼠子宫内膜间质细胞的活力、分化和迁移等生物学功能，并证实养血安胎方介导RAGE通路激活NF-κB信号促进EMT过程。此外通过观察RSA患者组织标本研究子宫内膜上皮细胞RAGE-NF-κB通路减弱EMT过程与反复流产的关系，并应用动物实验和细胞实验证明养血安胎方可通过RAGE-NF-κB通路影响EMT过程以达到治疗RSA的效果并阐释其作用机制，为养血安胎方治疗RSA提供科学依据。本研究为养血安胎方治疗RSA提供了一种新的EMT调控RSA的机制。