微电场对滋养细胞血管新生样功能和血管重铸活性的影响及其机制探讨

Extravillous trophoblast cells (EVTs) to invade uterine spiral artery,acquire endothelial phenotype and remodel the vessel are essential for the normal pregnancy,and insufficient remodeling will lead to severe pregnancy outcomes. We have recently demonstrated that applied small EFs could induce directed movement, elongation and enhanced expression of VEGF of EVTs, which is similar to EF-induced pre-angiogenic response of endothelial cells. We also found that EF could stimulate MMP-2 expression in the cells. Therefore we hypothesize that EF could promote angiogenic features and remodeling activity of trophoblast cells, and the mechanism might be related to activation of VEGF/VEGFR signalling pathway of the cells. Therefore,the present study is to measure tube formation of the cells cultured on matrigel, study VEGFR2 expression and activity and activity of its down-stream molecules, actin filament arrangement and angiogenic gene expression of trophoblast cells under EF stinulation.In addition, we also plan to study invasive feature of trophobast cells stimulated by EF into endothelial monolayer. No such studies have yet been reported. This study has potential significance not only for our understanding the role of EFs-induced placeental blood supply and related mechanism of it, but also has therapeutic potential to apply EFs preventing ischemia complications in placenta.

绒毛外滋养细胞（EVTs）侵入子宫螺旋动脉﹑获得内皮样表型并重铸血管为正常妊娠所必须，重塑不足则会导致严重的产科结局。我们近年发现，微电场(EF)可刺激EVTs定向运动和定向排列并分泌VEGF,与EF刺激内皮细胞血管新生反应相似；还发现EF可刺激EVTs的 MMP2表达增加。我们设想EF能促进EVTs血管新生样功能和血管重铸活性，其机制与VEGF信号通路的活化有关。为此，我们拟分析EF对matrigel培养滋养细胞管状结构形成的影响，测定EF刺激细胞VEGF受体2的表达和活化﹑胞内由VEGFR介导的血管新生信号分子的活化及actin胞内分布和血管新生相关的基因表达，并测定EF刺激EVTs对血管内皮细胞的侵袭作用。迄今尚未见EF对滋养细胞血管新生样功能和血管重铸活性影响及其机制的研究报道。本研究不仅对探讨EF改善胎盘血液供应功能具有重要理论意义，而且对应用EF防治胎盘缺血性疾病有应用价值。

绒毛外滋养细胞（EVTs）侵入子宫螺旋动脉﹑获得内皮样表型并重铸血管为正常妊娠所必须，重塑不足则会导致严重的产科结局。我们近年发现，微电场(EF)可刺激EVTs定向运动和定向排列并分泌VEGF,与EF刺激内皮细胞血管新生反应相似；还发现EF可刺激EVTs的MMP2表达增加。我们设想EF能促进EVTs血管新生样功能和血管重铸活性，其机制与VEGF信号通路的活化有关。为此，我们分析了EF对matrigel培养滋养细胞管状结构形成的影响，测定了EF刺激细胞VEGF受体2的表达和活化﹑胞内由VEGFR介导的血管新生信号分子的活化及actin胞内分布和血管新生相关的基因表达，并测定了EF刺激EVTs对血管内皮细胞的侵袭作用。. 本研究主要结果显示：微电场刺激不仅促进滋养细胞定向迁移和定向排列等血管新生样反应，也促进滋养细胞血管样结构的形成、生长密度及生长速度；微电场作为电信号使滋养细胞VEGF/VEGFR2信号通路激活，胞内重要信号分子P42/44 MAPK、AKT、FAK、Paxillin、eNOS及F-actin等发生活性的增加或分布的改变；微电场影响滋养细胞迁移、增殖及血管新生等相关分子的表达和基因表达谱的改变；微电场刺激能促进滋养细胞侵入血管内皮细胞的活性。. 本研究不仅对微电场作为一种可能的生理性信号影响胎盘血管新生/重铸功能及其机理的探索具有重要理论意义，而且对应用微电场促进胎盘血管新生防治胎盘缺血性疾病如流产、子痫前期和胎儿宫内生长受限等的发生具有实际意义和应用前景。.