VEGF 165基因转染HUVEC促进大鼠血管生成的实验研究

VEGF 165 Gene Transfected HUVEC Promotes Angiogenesis:an in vivo Study in Rats

目的:探讨VEGF 165基因转染脐静脉内皮细胞(HUVEC)移植在体内促进血管生成的能力。方法:常规复苏HUVEC后体外培养,经缺陷性腺病毒介导VEGF 165基因转染HUVEC。大鼠背部建立缺血皮瓣模型,实验组(10只),将VEGF 165基因转染的HUVEC移植于皮瓣局部;对照组(10只),移植未转染基因的HUVEC;空白组(10只)注射等量生理盐水。分别检测皮瓣存活率、外周血血清中VEGF浓度,行HE染色及CD31的免疫组化,并计算各组皮瓣微血管密度(microvessel density,MVD)。结果:不同时期,实验组外周血中VEGF浓度明显高于其他两组(P<0.05);对照组和空白组各个时期血清中VEGF浓度无明显变化(P>0.05)。皮瓣术后11 d,各组皮瓣存活率分别为实验组(72.5±4.7)%、对照组(45.8±3.6)%、空白组(32.4±1.7)%,3组皮瓣存活率比较差异有统计学意义(P<0.05)。HE染色及CD31免疫组化染色结果显示,实验组中毛细血管密度远大于其他两组(P<0.05)。结论:VEGF 165基因转染的HUVEC,移植在体内,具有显著地促进血管生成能力。

Objective:The aim of the study was to assess the angiogenesis potential of vascular endothelial growth factor(VEGF)165 gene transfected human umbilical vein endothelial cells(HUVEC)in vivo.Methods:Cultivating HUVEC after recovering in vitro,and then transfected with VEGF 165 gene to HUVEC through adenovirus.Ischemic skin flap model in back of Sprague?鄄Dawley rats were established,The experimental group(n=10)was transplanted HUVEC transfected with VEGF 165 gene.The control group(n=10)was transplanted HUVEC without VEGF 165 gene.The blank group(n=10)was injected with only NS.Detecting the VEGF density in peripheral blood serum with ELISA.Assessing the flap survival rate and taking specimen for pathologic examination and immunohistochemistry examination of CD31 to count microvessel density(MVD).Results:The VEGF density in experimental group always exceeds the other groups in any time point(P<0.05).The VEGF density between the other two groups have no significant difference in any time points(P>0.05).The comparison of flap survival rate between three groups demonstrated significant difference(P<0.05).The results of HE staining and immunohistochemistry of CD31 proved that,the MVD in experimental group was more than other groups.Conclusion:Transplanted HUVEC which were transfected with VEGF 165 gene have dramatically ability of promoting angiogenesis in vivo.