血管内皮生长因子165转染可促进脂肪间充质干细胞增殖

Vascular endothelial growth factor 165 transfection promotes proliferation of adipose-derived mesenchymal stem cells

背景:血管内皮生长因子是目前最重要的促血管新生因子之一,在脂肪移植中可促进移植物的血运重建,提高成活率。目的:探讨血管内皮生长因子165基因转染促进脂肪间充质干细胞增殖的作用。方法:利用目的片段重组血管内皮生长因子165基因入腺病毒p Ad Easy-1系统,包装病毒并测定滴度,同理包装空病毒。将包装成功的两种病毒液分别以100的最佳感染复数转染入脂肪间充质干细胞内,以未转染细胞为空白组。采用RT-PCR和Western blot法检测各组转染细胞中血管内皮生长因子165 m RNA和蛋白的表达,MTT法检测各组细胞的增殖情况。结果与结论:实验组血管内皮生长因子165 m RNA和蛋白表达量高于对照组和空白组(P<0.05);实验组转染脂肪间充质干细胞后分裂增殖程度显著增加,与对照组和空白组比较差异有显著性意义(P<0.05),结果表明腺病毒介导的血管内皮生长因子165基因转染脂肪间充质干细胞后可以持续表达目的蛋白,同时也能促进脂肪间充质干细胞的显著增殖。

BACKGROUND： Vascular endothelial growth factor is the most important pro-angiogenic factor, which can promote revascularization and survival of fat grafts during fat transplantation. OBJECTIVE： To investigate the effect of vascular endothelial growth factor 165 transfection on the proliferation of adipose-derived mesenchymal stem cells. METHODS： Recombinant vascular endothelial growth factor 165 m RNA fragment was transmitted into adenovirus p Ad Easy-1 systems that were packaged to measure viral titer（experimental group）. Empty adenovirus was also packaged as control group. Two kinds of packaged adenovirus solution were transferred into adipose-derived mesenchymal stem cells at the best multiplicity of infection=100. Cells with no transfection served as blank group. RT-PCR and western blot methods were used to detect the expression of vascular endothelial growth factor 165 at m RNA and protein levels; MTT method was adopted to detect cell proliferation. RESULTS AND CONCLUSION： The expressions of vascular endothelial growth factor 165 m RNA and protein were significantly higher in the experimental group than the control and blank groups（P〈0.05）. The division and proliferation of transfected adipose-derived mesenchymal stem cells were increased significantly in the experimental group, which was significantly different from the control and blank groups（P〈0.05）. These finding indicate that vascular endothelial growth factor 165 transfection cannot only sustain the target protein expression of adipose- derived mesenchymal stem cells, but also promote the proliferation of adipose-derived mesenchymal stem cells remarkably.