基于miRNAs研究不同频率电针调控LIF/JAK/STAT3提高超促排卵围着床期子宫内膜容受性的作用机制

Dysfunction of cell junction remodeling in endometrial epithelial cells is one of the defining pathological features of decreasing of endometrium receptivity(DER). LIF/JAK/STAT3 signaling pathway involves in the process of cell junction remodeling mediated by STAT3 pophosphorylation, and is associated with the miRNAs levels. Abnormal miRNAs levels in endometrium during controlled ovarian hyperstimulation cycles can impact LIF/JAK/STAT3 signaling pathway, and then inhibits cell junction remodeling, leading to a decreased endometrium receptivity and repeated blastocyst implantation failure. Focusing on correcting abnormal miRNAs , we will adopt acupuncture and molecular biological technologies in a COH-like pathology rat model, on the basis of previous studies on the effects of acupuncture for decreasing of endometrium receptivity during controlled ovarian hyperstimulation cycles, to study the underlying mechanisms of acupuncture on enhancing cell junction remodeling in endometrial epithelial cells and ameliorating endometrium receptivity , and observe the effect differences of electroacupuncture with different frequencies. The mechanisms would greatly reveal the understanding of acupuncture effect on DER and provide scientific experimental evidence for prompting acupuncture treatment to treat DER.

子宫内膜腔上皮细胞间连接重塑受限是内膜容受性低下的主要病理改变之一。宫腔上皮细胞中LIF/JAK/STAT3 信号转导通路参与STAT3磷酸化介导的细胞间连接重塑机制，且与miRNAs水平相关。COH周期子宫内膜miRNAs水平紊乱可直接影响LIF/JAK/STAT3通路，从而抑制宫内膜腔上皮细胞间连接重塑，导致内膜容受性低下，造成反复胚胎着床失败。本项目拟在前期防治COH内膜容受性低下的研究基础上，以COH模型鼠为对象，予针刺干预，应用相关分子生物学技术，从调控LIF/JAK/STAT3转导通路相关表观遗传修饰的角度，研究电针通过调控COH周期miRNAs水平以增强宫腔上皮细胞间连接重塑、改善内膜容受性的作用机制，并观察不同频率电针的效应差异，为针灸防治COH周期内膜容受性低下提供科学的实验依据。

本研究以超促排卵模型大鼠为研究对象，以高、低频(50Hz、2Hz)电针为干预手段，从子宫内膜腔上皮细胞间连接重塑功能受限的病理特点入手，结合表观遗传修饰在RIF-IVF 内膜容受性低下发病中调控的重要性，以LIF表达及STAT3蛋白磷酸化过程相关miRNA为切入点，运用分子生物学技术从不同角度研究超促排卵模型鼠着床窗口期子宫内膜LIF表达及STAT3磷酸化过程的表观遗传修饰在着床窗口期对内膜容受性的影响，分析高、低频电针对子宫内膜LIF/JAK/STAT信号转导通路及子宫内膜腔上皮中E-cadherin、β-catenin、Claudin1 三类相关蛋白表达的影响。研究结果显示电针，特别是高频电针，可通过下调COH模型大鼠围着床期子宫内膜miR-223-3p，继而串化式调控LIF/JAK/STAT信号转导通路及下游细胞连接蛋白的表达，以减少胚泡着床的机械屏障，提高内膜容受能力，从而促进胚胎黏附及种植。此研究为优化临床电针频率选择、应用和推广电针治疗COH相关内膜容受性低下提供了科学的实验依据。