腺病毒转染血管内皮生长因子基因对感染大鼠骨髓间充质干细胞的影响

Effect of transfecting VEGF gene with AdCMV vector on rat bone marrow mesenchymal stem cells

目的体外观察和检测腺病毒AdCMV转染血管内皮生长因子(VEGF)基因对大鼠骨髓间充质干细胞(BMSCs)增殖的影响。方法培养大鼠BMSCs,用AdCMV-增强型绿色荧光蛋白(eGFP)、AdCMV-VEGF感染大鼠BMSCs,MTT检测AdCMV-VEGF对大鼠BMSCs增殖的影响,Western blot检测不同浓度的AdCMV-VEGF感染大鼠BMSCs后蛋白的表达量和BMSCs组、AdCMV-eGFP组、AdCMV-VEGF组细胞的VEGF蛋白表达量的差异。结果成功培养了BMSCs。与BMSCs组和AdCMV-eGFP组比较,AdCMV-VEGF组的VEGF蛋白表达较多。结论成功获得大鼠BMSCs,且AdCMV-VEGF成功转染大鼠BMSCs,为缺血性脑血管疾病治疗提供有用的参考。

Objective To study the effect of transfecting VEGF gene with AdCMV vector on rat bone marrow mesenchymal stem cells（BMSCs） in vitro. Methods The rat BMSCs were cultured in vitro and infected with AdCMV-eGFP and AdCMV-VEGF. The effect of transfecting VEGF on the proliferation of rat BMSCs was evaluated by MTT. The expression of VEGF gene in BMSCs transfected with different concentrations of AdCMV vector was detected by Western blot. The difference of VEGF gene expression among groups of AdCMV-eGFP, AdCMV-VEGF and untransfected cells were detected by Western blot as well. Results The BMSCs were cultured successfully. The expression of VEGF protein was higher in AdCMV-VEGF-BMSCs than that in groups of BMSCs and AdCMV-eGFP-BMSCs. Conclusion The rat BMSCs have been successfully cultured and transfected by AdCMV-VEGF, which may provide an useful reference in the treatment of cerebral vascular disease.