miRNA对子宫内膜蠕动波的调控及其机制研究

Uterine peristalsis plays an important role in embryo implantation and could be a non-invasive criterion to predict endometrial receptivity. However, the mechanism of uterine peristalsis regulation is unclear.miRNAs take part in various kinds of endometrial physiological activities and implantation of embryo.While few miRNAs were known about uterine peristalsis regulation. Thus, studies are designed to find out the related miRNA and target mRNA by miRNA-mRNA microarray conjoint analysis. Influence of miRNA and its target gene expression changes to intracellular calcium ion concentration and movement of endometrial stromal cells and smooth muscle cells beneath endometrium are analyzed. The regulation role of target gene is confirmed in a cell co-culture model to clarify the mechanism of uterine peristalsis . In conclusion, this study connects uterus specifically expressed miRNA with uterine peristalsis and endometrial receptivity, has important implications for clarifying the mechanism of uterine peristalsis and its influence on embryo implantation, and lays the foundation for the treatment of abnormal uterine peristalsis.

子宫内膜蠕动波是一种无创性评估子宫内膜容受性的重要指标，在胚胎着床中起关键作用，但调控子宫内膜蠕动波的机制不清。miRNAs参与调节子宫内膜的各种生理活动及胚胎的植入，但目前所知的参与子宫内膜蠕动波调控的miRNA甚少。为筛选调控子宫内膜波状运动的miRNA，我们拟采用miRNA-mRNA芯片联合分析，筛选并验证调控子宫内膜蠕动波的miRNA及其靶mRNA基因；分析miRNA及其靶基因表达改变对子宫内膜间质细胞及内膜下肌层平滑肌细胞的细胞内钙离子浓度和运动的影响；并在细胞共培养模型中验证靶基因对子宫内膜蠕动波的调控功能。本研究将子宫特异表达的miRNA与子宫内膜蠕动波、子宫内膜容受性相联系，对阐明子宫蠕动波的发生机制及其对胚胎着床的影响具有重要意义，为异常子宫内膜蠕动波的临床治疗奠定基础。

子宫内膜蠕动波是一种无创性评估子宫内膜容受性的重要指标，在胚胎着床中起关键作用，但调控子宫内膜蠕动波的机制不清。miRNAs参与调节子宫内膜的各种生理活动及胚胎的植入，但目前并无参与子宫内膜蠕动波调控的miRNA的研究。通过对miRNA芯片以及RT-qPCR技术筛查高频或低频子宫内膜蠕动波患者内膜的差异表达miRNA，原位杂交技术定位miRNA的分布，生物信息学结合文献资料预测miRNA的靶基因，免疫组化定位与半定量分析预测靶基因在内膜的表达，双荧光素酶报告基因实验验证miRNA的靶基因，再通过子宫内膜间质细胞与子宫平滑肌细胞的共培养及凝胶收缩实验分析子宫内膜miRNA如何影响子宫内膜蠕动波，再通过WB、ELISA等技术阐述其具体机制。结果发现miR-29c-3p在高频组子宫内膜蠕动波患者中的表达比低频组子宫内膜蠕动波患者更低，主要定位在人子宫内膜间质细胞的胞质中。子宫内膜间质细胞miR-29c-3p能靶向结合并下调OXTR的表达。OXTR既表达于人子宫内膜腺上皮细胞的胞质胞核也表达于子宫内膜间质细胞的胞质胞核，且OXTR在高频组子宫内膜表达比低频组更高（0.056±0.0059 V.S.0.036±0.0045，P<0.01）。将子宫内膜间质细胞与子宫平滑肌细胞共培养于胶原蛋白凝胶中，发现miR-29c-3p上调的子宫内膜间质细胞与平滑肌细胞共培养组的凝胶面积较对照组大（P<0.05），miR-29c-3p下调的子宫内膜间质细胞与平滑肌共培养组的凝胶面积较对照组小 (P<0.05)。仅将转染了miR-29c-3p mimic或inhibitor的子宫内膜间质细胞种植于胶原蛋白凝胶中与相应的对照组比，面积无明显改变（P>0.05）。miR-29c-3p上调的子宫内膜间质细胞能减弱AKR1C3表达以及PGF2α的产生和释放，miR-29c-3p下调时则呈相反的趋势。总之，子宫内膜间质细胞miR-29c-3p能通过降低OXTR、AKR1C3的表达、影响PGF2α的分泌，影响子宫内膜下平滑肌组织的收缩，从而参与子宫内膜蠕动波的调控。该研究表明miRNA参与了子宫内膜蠕动波的调控及其主要机制，能为异常子宫内膜蠕动波的治疗提供理论依据及实验基础，从而改善患者的子宫内膜容受性改善妊娠结局。