缺氧/miR-21/HGF-sMet信号系统在重度子痫前期发病过程中的调控研究

PE is a pregnancy-specific syndrome characterized by maternal hypertension and proteinuria occurring after the 20th week of gestation. By far the etiology and pathogenesis of preeclampsia has not been well understood.Our preliminary data showed that the expression of miR-21 was upregulated in severe preeclampsia placenta by 2-fold compared with that in control group.In this project,we mainly aim to: 1)compare miR-21 expression in placenta and plasma between severe preeclampsia and normal pregnancy by real time PCR and in situ hybridization, and evaluate the predictive values of miR-21 for severe preeclampsia；2)study the effects of miR-21 on trophoblast and endothelial vascular function by testing the ability of cell outgrowth,invasion, tube formation of HUVEC on Matrigel in vitro and the Matrigel plug assay in vivo；3)identify the interaction of miR-21 with HGF/c-Met angiogenic system by Dual-Luciferase Reporter Assay System,and to detect whether miR-21 is responsible for the shedding of c-Met via upregulating matrix metalloproteinase and ADAMs activities；4)to create the adenovirus-mediated miR-21 overexpression rat model,and to evaluate whether miR-21 contribute to severe preeclampsia by regulating HGF/c-Met angiogenic system；5)to determine whether hypoxia can regulate miR-21 expression. This study may give new insight for the etiology and pathogenesis of preeclampsia, and to provide a new potential bio-marker for early prediction of risk for developing preeclampsia,and a novel therapeutic target for preeclampsia.

子痫前期是妊娠并发症之一，发病机理仍不清楚。前期工作证实miR-21 在重度子痫前期患者胎盘中水平升高，为对照组水平2倍。本项目运用荧光定量PCR技术等检测miR-21在重度子痫前期患者和正常孕妇胎盘、血浆中的差异水平，ROC 曲线评价子痫前期发生风险；通过滋养层细胞增殖、侵润实验，HUVEC 管状结构形成及在体Matrigel 栓子试验，研究miR-21调控滋养层细胞和血管内皮细胞功能的作用；荧光报告基因系统验证miR-21 与HGF的相互作用,研究miR-21是否上调MMPs和ADAMs而促进soluble Met(sMet)产生释放；建立过表达miR-21的大鼠模型，研究miR-21调节HGF/c-Met及sMet在重度子痫前期发病中的作用机制；研究缺氧对miR-21 水平的调节与重度子痫前期发病的相关性。为子痫前期病因学研究提供新的数据，为子痫前期早期诊断、治疗提供新的指标和靶点。