论文部分内容阅读
目的 :探讨外源性人VEGF16 5基因在内皮细胞与心肌细胞内表达的可行性。方法 :构建了 (vascularendothelium growthfactor,VEGF)与增强型绿色荧光蛋白 (enhancedgreenfluores cent protein ,EGFP )基因共表达载体 pIRES2 EGFP VEGF16 5 ,以脂质体法转染内皮细胞和心肌细胞 ,采用荧光显微镜检测内皮细胞与心肌细胞中EGFP的表达 ,同时利用免疫组织化学方法检测VEGF的表达。结果 :成功地构建了真核表达载体 pIRES2 EGFP hVEGF16 5 ,采用脂质体法转染内皮细胞与心肌细胞后 ,经荧光显微镜观察 ,可见细胞内有EGFP的表达 ,同时经免疫组化证实有VEGF的表达。结论 :采用脂质体法可以成功地将外源性VEGF16 5基因转染到内皮细胞与心肌细胞中 ,并进行表达。本研究为今后利用VEGF基因治疗心肌缺血等疾病提供了实验基础。
Objective: To investigate the feasibility of exogenous human VEGF16 5 gene expression in endothelial cells and cardiomyocytes. Methods: The co-expression vector of VEGF and enhanced green fluorescent protein (EGFP) gene, pIRES2 EGFP VEGF165, was constructed and transfected into endothelial cells and cardiomyocytes by lipofectamine 2000. Endothelial EGFP expression in cells and cardiomyocytes, meanwhile, the expression of VEGF was detected by immunohistochemistry. Results: The eukaryotic expression vector pIRES2 EGFP hVEGF16 was constructed successfully. After transfection of endothelial cells and cardiomyocytes by lipofectamine, the expression of EGFP in the cells was observed by fluorescence microscope. VEGF was confirmed by immunohistochemistry expression. Conclusion: The exogenous VEGF165 gene can be successfully transfected into endothelial cells and cardiomyocytes by liposome method and expressed. This study provides the experimental basis for the future use of VEGF gene therapy for diseases such as myocardial ischemia.