微电场刺激三维培养血管内皮细胞血管新生及其机制的研究

Angiogenesis is involved in a variety of physiological and pathological process. Physiological small electric fields (EFs) exist in and around the vasculature in the regions of active angiogenesis. We recently demonstrated that applied small EFs could stimulate pre-angiogenic response of endothelial cells in 2D culture, and the effect was due to activation of VEGF/VEGF receptors (VEGFRs) signaling pathway. 3D culture procedure of angiogenesis appears to place the endothelial cells in a more physiological state mimicking in vivo conditions.The present study is to quantify EFs-induced angiogenic responses in 3D models as the tube formation, direction of tube growth, tube density and growth rate, and to explore applied small EFs inducing distribution and /or expression of membrane receptors and introcellular molecules relating to angiogenesis, therefore gain insight into the molecular mechanism underlying the EFs-induced angiogenic responses in endothelial cells. No such studies have yet been reported. This study has potential significance not only for our understanding the role of EFs-induced angiogenic responses in endothelial cells and the related mechanism of it, but also has therapeutic potential by enhancing local blood supply in ischemia diseases as well as inhibiting EFs to inhibit tumor growth, diabetic retinopathy, and vascular involvement in rheumatic diseases.

血管新生参与机体重要的生理和病理过程。生理性微电场在活体血管新生活跃部位广泛存在。我们发现微电场可刺激2D培养血管内皮细胞血管新生前期样反应，并与VEGF/VEGFR信号通路有关。鉴于3D培养血管内皮细胞血管新生模型更接近活体状态，本研究拟定量测定微电场诱导3D培养血管内皮细胞血管样结构的形成、生长方向、密度及生长速度，并探索微电场诱发血管内皮细胞膜和胞内与血管新生有关受体和信号传递分子的分布和表达情况，以了解微电场诱导血管新生反应的分子机理。研究生理性微电场对三维培养血管内皮细胞血管新生作用的影响及其相关信号分子的作用机理，国内外尚未见报道。本研究不仅对探讨微电信号影响内皮细胞血管新生功能及其机理有重要理论意义，而且对应用微电场刺激改善缺血性疾病的局部血液供给，以及抑制微电场对血管新生的作用以抑制肿瘤生长、糖尿病视网膜病变及类风湿等炎症性血管疾病有应用前景。

血管新生参与机体重要的生理和病理过程。生理性微电场在活体血管新生活跃部位广泛存在。我们发现微电场可刺激2D培养血管内皮细胞血管新生前期样反应，并与VEGF/VEGFR信号通路有关。鉴于3D培养血管内皮细胞血管新生模型更接近活体状态，本项目分析了微电场诱导3D培养血管内皮细胞管样结构形成的作用，测定了微电场诱发血管内皮细胞与血管新生有关VEGF/VEGFR2及下游信号通路分子的表达活化情况；在单层细胞培养模型上进一步观察了微电场对血管内皮细胞迁移、存活和细胞增殖等血管新生相关的信号传递分子的影响以及相关关键基因和基因表达谱等的改变，并在体外器官水平初步观察了微电场刺激小鼠主动脉环新生微血管的生长情况。.本研究主要结果显示：微电场不仅影响二维培养血管内皮细胞血管新生前期样反应，也影响三维培养血管内皮细胞血管样结构的形成、密度、生长速度及生长方向；微电场作为电信号使血管内皮细胞VEGF/VEGFR信号通路激活，胞内重要信号分子发生表达和/或活性的改变；微电场影响血管内皮细胞血管新生相关分子的表达和基因表达谱的改变。.本研究不仅对探讨微电信号影响内皮细胞血管新生功能及其机理有重要理论意义，而且对应用微电场刺激改善缺血性疾病的局部血液供给，以及抑制微电场对血管新生的作用以抑制肿瘤生长、糖尿病视网膜病变及类风湿等炎症性血管疾病有应用前景。