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目的 构建血管内皮生长因子(vascular endothelial growth factor,VEGF) 165基因的腺病毒表达载体,并观察其感染血管内皮细胞后目的基因的表达及对皮瓣存活的影响.方法 采用复制缺陷型重组腺病毒载体介导的VEGF165基因(Ad-VEGF165)感染原代培养血管内皮细胞和大鼠背部随意皮瓣组织,分别从体外和体内实验两方面研究Ad-VEGF165感染对靶细胞或/和组织生物学特征和血管生成的影响,实时定量聚合酶链反应(qPCR)和Western印迹分析法(Western Blot)检测腺病毒介导的外源性VEGF165基因在血管内皮细胞中的转录和表达,观察其增强皮瓣存活的潜力.结果 Ad-VEGF165转染后,血管内皮细胞中的VEGF165含量较对照组和腺病毒空载体感染组均明显升高(P＜0.05);Ad-VEGF165组、腺病毒空载体组和对照组比较,皮瓣组织中的VEGF165表达差异显著(P＜0.05),血管生长更旺盛.结论 Ad-VEGF165感染皮瓣组织后,能显著增加VEGF165蛋白表达水平,促进血管生成,为进一步开展VEGF165基因提高皮瓣移植成活率的临床研究奠定了实验基础.“,”Objective To construct a recombinant adenoviral vector carrying and expressing human vascular endothelial cell growth factor 165 (Ad-VEGF165) and examine the expression of VEGF165 in Ad-VEGF165-infected vascular endothelial cells and its effect on flap survival.Methods Human umbilical vein endothelialcells (HUVECs) in primary culture and random skin flaps were transfected with VEGF165 gene encoded in the replication-deficient recombinant adenovirus.The effects of Ad-VEGF165 on the biological features and angiogenesis of transfected cells or tissues were investigated in both the in vitro and in vivo systems.Adenovirus-mediated transcription and expression of exogenous VEGF165 gene in HUVECs was examined by qPCR and Western Blot.The effect of adenovirus-mediated VEGF gene therapy on the survival of random skin flap in rats was evaluated.Results Compared to control HUVECs and blank adenovirus transfected HUVECs, Ad-VEGF165-infected HUVECs had significantly higher levels of VEGF165 transcription and expression (P ＜ 0.05).Same increase of VEGF165 expression was seen in the flaps with subsequent increased angiogenesis.Conclusion VEGF expression is significantly enhanced in flap tissues that are transfected with Ad-VEGF165.This stimulates angiogenesis in the tissues and improves flap survival.These experiments have laid the foundation for further research on VEGF165 gene therapy.