骨形态发生蛋白-2与血管内皮生长因子双基因质粒转染鼠骨髓间充质干细胞的研究

Bone marrow mesenchymal stem cells transfected by a dual-gene coexpression plasmid of bone morphogenetic protein-2 and vascular endothelial growth factor

目的 利用骨形态发生蛋白-2（BMP-2）、血管内皮生长因子（VEGF）基因构建双基因质粒,转染鼠骨髓间充质干细胞,通过基因表达,促进成骨、成血管能力.方法 构建BMP-2、VEGF基因及双基因质粒,分离培养鼠骨髓间充质干细胞（BMSCs）,进行质粒转染,在激发波长488 nm、发射波长507 nm荧光显微镜下观察并计算转染率,分别于1、4、7、10、15、21 d采用酶联免疫吸附试验（ELISA）检测转染了各组质粒的BMSCs培养液中BMP-2、VEGF蛋白的表达.结果 转染结果显示,BMP-2组为38.2％,VEGF组为40.1％、双基因质粒组为37.2％.结果显示各组均有目的蛋白的分泌,BMSCs及空载体组在BMP-2、VEGF的表达上差异无统计学意义（P＞0.05）,其余各组释放量比较差异有统计学意义（P＜0.05）,双基因质粒组的两种蛋白分泌量大于其他各组.结论 双基因质粒的表达可以互相促进,起到级联放大作用.

Objective The bone marrow mesenchymal stem cells （BMSCs） transfected by a dual-gene plasmid of bone morphogenetic protein-2 （BMP-2） and vascular endothelial growth factor （VEGF） gene.Through the gene expression,promote the angiogenesis and osteogenic activity.Methods We construct the BMP,VEGF and dual-gene plasmid.Separation of rat bone marrow mesenchymal stem cells （BMSCs）,the cells were transfected by the plasmid,At excitation wavelength 488 nm and emission wavelength of 507 nm,under the fluorescence observed the ratio of the transfection,the gene exoression.Results The ratio of BMP-2 group is 38.2％,the group of VEGF is 40.1％,the group of dual-gene is 37.2％.The results shows that every group secret the protein.The groups of BMSCs transfected by plasmids have significant difference.The protein of dual-gene group is more than the others groups.Conclusion The expression of dual-gene plasmid have the ability of mutual promotion.