VEGF转染VEC促进移植皮瓣血管化的实验研究

目的探讨VEGF165基因转染血管内皮细胞促进移植皮瓣血管化的可行性,提高皮瓣存活率。方法利用脂质体介导pIRES2-EGFP/VEGF165质粒体外转染VEC。27只SD大鼠随机分为3组,转染组和内皮细胞组大鼠缺血皮瓣内分别加入1ml浓度为1×106个/亳升的含有转染质粒pIRES2-EGFP/VEGF165血管内皮细胞和未转染正常血管内皮细胞的DMEM培养基,对照组注射相同量DMEM培养基。皮瓣早期断蒂,观察皮瓣的皮瓣存活率、HE染色和CD34免疫组化染色观察血管生成情况,计算微血管密度。结果VEGF165基因转染VEC、单纯的VECC移植大鼠缺血皮瓣后,与对照组的皮瓣存活率分别为98.6%±3.9%、83.2%±3.5%、42.5%±2.1%,差异有统计学意义(p<0.05)。术后第7d时三组皮瓣中点处的毛细血管密度分别为(60.4±6.5)个/毫米2、(58.2±6.3)个/毫米2、(17.8±1.7)个/毫米2;第14d时毛细血管密度分别为(91.9±7.1)个/毫米2、(74.4±6.8)个/毫米2、(28.6±3.7)个/毫米2。结论VEGF165转染血管内皮细胞可以可促进缺血皮瓣的血管新生,提高存活率。

TRANSPLANTATION OF VEGF -TRANSFECTED VASCULAR ENDOTHELIAL CELLS FOR IMPROVING THE VASCULARIZATION OF FLAP

Objective To study the feasibility of VEC transfeeted with VEGF165 for improving the vascularization of flap and the survivial rate. Method A eukaryotic expression vector pIRES2 - EGFP/VEGF165 was transfected by tiposome DOTAP into VEC. Twenty - seven male SD rats were randomly divided into three groups, named group A, B and C respectively. The rats in group A received DMEM culture medium lml （density is 1 × 10^6/ml ） including VEC transfected by pIRES2 -EGFP/VEGF165 plasmid,The rats in group B underwent VEC, while those in group C were transplanted only DMEM culture medium lml. The survivial rate of flap was observed. HE and immunohistochemical staining was used to detect the vascularization and calculate the microvessel density （MVD）. Result The flap survival rates of VEGF165 gene transfection group, VEC group, and the control group were 98.6% ± 3.9%, 83.2% ± 3.5 %, 42.5 % ± 2.1% respectively, with significant differences （ P 〈 0.05 ）. 7 days after operation, photo analysis shows that microvessels density of VEGF165 gene transfection group, VEC group, and the control group were 60.4 ±6.5/mm2, 58.2 ±6.3/mm2, 17.8 ± 1.7/mm2 respectively. 14days, MVD of 3 groups were 91.9 ± 7.1/ mm2, 74.4 ± 6.8/mm2, 28.6 ± 3.7/mm2 respectively. Conclusion The VEGF165 - transfected VEC can effectively stimulate the angiogenesis in flap in the early stage and further improve the survival rate.