人血管内皮细胞生长因子165基因转染大鼠内皮祖细胞的实验研究

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）及对其影响。方法体外分离、培养、鉴定大鼠骨髓EPCs,ELISA检测转基因EPCs表达VEGF蛋白,MTT法检测对其增殖的影响。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,注射转基因EPCs的大鼠皮下局部血管增加。结论hVEGF165基因可成功转染EPCs,并且具有血管内皮增殖刺激活性,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。

Human VEGF 165 gene transfection into rat endothelial progenitor cells

ObjectiveTo explore the feasibility of human VEGF165 gene transfection into endothelial progenitor cells(EPCs),as well as its influence on EPCs proliferation.MethodsEPCs were isolated from rat bone marrow,cultured in vitro,identified by FITC-UEA-I and Dil-acLDL staining.After transfection,the expression of VEGF was detected by ELISA.The influence on EPCs proliferation induced by gene transfection was studied by MTT methods.ResultsThe double FITC-UEA-I and Dil-ac-LDL fluorescence staining displayed differentiated EPCs.The expression of VEGF was found in the medium of EPCs transfected with pcDNA3hVEGF165.VEGF165 gene transfection could stimulate EPCs proliferation.ConclusionVEGF165 gene can be successfully transfected into EPCs,and the transduced EPCs enables to induce vascular growth.It offers a strong evidence for introducing the VEGF165 gene in EPCs in vivo.