VEGF基因转染血管内皮祖细胞的实验研究

目的:探讨血管内皮生长因子（VEGF）基因转染血管内皮祖细胞（EPCs）的方法及效果。方法:取生长活跃的EPCs，分别转染不同浓度的携带VEGF-165基因的腺病毒质粒（Adv-GFP-VEGF165）（转染组）及空质粒（Adv-GFP）（空质粒组），并设空白对照组。荧光显微镜下观察转染效果，MTT法检测不同病毒滴度时细胞增殖情况；ABC-ELISA法检测上清液中VEGF蛋白表达情况。结果:转染组镜下观察到大部分细胞呈现绿色荧光。病毒滴度为1∶100的转染会导致细胞生长停滞，并造成细胞损害； 1∶50的转染后对细胞增殖无明显影响。ELISA检测证实转染组上清液中VEGF蛋白浓度明显高于空质粒组及空白对照组（P<0.01）。结论:以腺病毒为载体的VEGF基因转染EPCs是可行的，1∶0是合适的转染比率，转染后上清液中VEGF蛋白表达明显增加。

Experimental study of endothelial progenitor cells transfected with VEGF165 gene

Abstract:Objective:To investigate the method and effect of endothelial progenitor cells (EPCs) transfected with VEGF165 gene.Methods :The cultivated EPCs were transfected with recombined adenovirus in which VEGF165 gene (Adv-GFP-VEGF165 group) or only Adv-GFP (Adv group) was incorporated,then the transfection effect was observed by immunofluorescence. And a blank control group was set up. Observation of the proliferation potential of cells was done by MTT method. The VEGF gene expression was observed by detecting the concentration of VEGF protein in supernatant with ABC-ELISA method.Results:Green fluorescence was observed in almost all of the cells after transfection with Adv-GFP-VEGF165, which indicated the success of the transfection. The EPCs proliferation was not affected after the transfection, which was confirmed by MTT method. The VEGF protein concentration in the supernatant of transfected group was higher than that of Adv group and control group.Conclusions: It is feasible for EPCs to be transfected with VEGF165 gene which was incorporated into adenovirus, and the suitable ratio was 1〖JP20〗∶〖JP〗50.The VEGF protein concentration in the supernatant liquid of endothelial progenitor cells transfected with VEGF increased significantly.