血管内皮生长因子基因修饰内皮祖细胞对内膜损伤血管修复的影响

Re-endothelialization of VEGF-transfected endothelial progenitor cells in rat artery intimal injury model

目的探讨人血管内皮细胞生长因子165(hVEGF165)基因转染骨髓源性内皮祖细胞(EPCs)及对EPCs增殖的影响,在体研究VEGF基因修饰EPCs对内膜损伤血管再内皮化的影响。方法体外分离、培养、鉴定EPCs,脂质体介导pcDNA3-hVEGF165质粒转染组、pcDNA3空质粒转染组、空白对照组。ELISA法检测EPCs表达VEGF蛋白,MTT法检测hVEGF165基因修饰对EPCs增殖的影响。基因修饰EPCs移植到内膜损伤血管模型中,观察基因修饰对EPCs修复内膜损伤的效应。结果FITC-UEA-I和DiI-acLDL荧光双染证实分化的EPCs,脂质体介导pcDNA3-hVEGF165质粒转染组EPCs培养基上清中表达VEGF,hVEGF165基因转染促进EPCs迁移、黏附和增殖。基因修饰EPCs促进损伤内膜修复。结论EPCs可以成功转染hVEGF165基因,并且促进EPCs的迁移、黏附和增殖,增强EPCs对损伤内膜修复的能力。

Objective To explore the proliferation of endothelial progenitor cells （EPCs） after the transfection of human VEGF165, and investigate the re-endothelialization of the transfected cells in artery intimal injury rats. Methods After isolated from rat bone marrow, EPCs were cultured and then identified by FITC- UEA-I and Dil-acLDL. EPCs cells were transfected with pcDNA3-hVEGF165 or pcDNA3 by DOTAP Liposome. After transfection, the expression of VEGF was detected by using ELISA. The EPCs proliferation was studied by MTT methods. VEGF-transfected EPCs were transplanted into experimental artery intimal injury rats and its effect on injured vascular re-endothelial was studied. Results Cells expressed double FITC-UEA-I and DilacLDL fluorescence were confirmed to EPCs. The medium of EPCs transfected with pcDNA3-hVEGF165 had expression of VEGF. VEGF165 gene transfer stimulated EPCs adhesion, migration and proliferation. Genemodified EPCs transplanted into rats, survived in intimal vascular injury site, repaired intimal vascular injury better than those in normal EPCs group. Conclusion VEGF165 gene is successfully transfected into EPCs, which can stimulate EPCs＇ migration, adherence and proliferation. VEGF-transfected EPCs have stronger intimal repair ability compared with non-transfected EPCs. VEGF-transfected EPCs repair injured vascular intima, restore normal vascular endothelial structure.