人VEGF 165基因转染大鼠内皮祖细胞及对其增殖的影响

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。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检测EPCs表达VEGF蛋白,MTT法检测hVEGF165基因修饰对EPCs增殖的影响。结果FITC-UEA-Ⅰ和DiI-ac-LDL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增殖。结论EPCs可以成功转染hVEGF165基因,并且可促进EPCs的增殖,为进一步研究hVEGF165基因修饰EPCs治疗血管内膜损伤性疾病提供实验依据。

Expression of human VEGF165 gene transfected into rat endothelial progenitor cells and effects on cells proliferation

Objectives To explore the feasibility of human VEGF165 gene transfer to endothelial progenitor cells(EPCs),and influencing on EPCs proliferation.Methods EPCs were isolated from rat bone marrow,cultured in vitro,identified by FITC-UEA-I and Dil-ac-LDL.EPCs transfected with pcDNA3-hVEGF165 by DOTAP Liposomal,EPCs transfected with pcDNA3 by DOTAP Liposomal and EPCs without transfection.After transfection the expression of VEGF was detected by using ELISA.The influence on EPCs proliferation induced by gene transfer was studied by MTT methods.Results Cells expressed double FITC-UEA-I and Dil-ac-LDL fluorescence were EPCs.The medium of EPCs transfected with pcDNA3-hVEGF165 had expression of VEGF.VEGF165 gene transfer could stimulate EPCs proliferation.Conclusion VEGF165 gene could be successfully transfected into EPCs,VEGF produced by gene thansfer can stimulate EPCs proliferation,it can give strong evidence for introducing the VEGF165 gene in EPCs in vivo.