复制缺陷型腺病毒载体介导的VEGF基因治疗大鼠缺血皮瓣的实验研究  

The effect of adenover-mediated VEGF gene on survival of a random skin flap in the rat

作　　者：王伟[1] 汪春兰[1] 赵宇[1] 曹东升[1] 王帮河[1] 丁浩[1] 王新宇[1] 朱华锋[1] 

机构地区：[1]安徽医科大学第一附属医院整形外科,合肥230032

出　　处：《安徽医科大学学报》2005年第4期302-304,共3页Acta Universitatis Medicinalis Anhui

摘　　要：目的探讨局部应用以复制缺陷型腺病毒为载体的血管内皮生长因子(Ad-VEGF165cDNA)基因治疗对大鼠随意缺血皮瓣生存的影响.方法选用30只SD大鼠,体重350～400 g,随机分成三组,在其背部形成8 cm×2 cm的全厚随意型皮瓣.于皮瓣远端真皮下分别注射Ad-VEGF165cDNA(10只目的基因组)、Ad-GFP(10只绿色荧光蛋白基因对照组)、PBS(10只磷酸盐缓冲液空白对照组).48 h后按原设计掀起皮瓣并原位缝合,术后7d,测量皮瓣的成活面积,并采集组织行免疫组化、组织学检查等.结果皮瓣成活率: Ad-VEGF165cDNA组为(74.382±1.338)%,Ad-GFP组为(52.246±1.663)%, PBS组为(52.871±2.890)%,Ad-VEGF165cDNA组与其他两组间差异有显著性(P<0.01),免疫组化显示基因转染组毛细血管周围VEGF沉积,其微血管密度明显高于对照组.结论局部皮下注射Ad-VEGF165cDNA,可诱导局部VEGF蛋白表达,促进新生血管的形成,提高缺血皮瓣的成活面积,是一种简单有效的基因治疗方法.Objective To investigate the effect of adenovirus-mediated gene therapy with vascular endothelial growth factor （VEGF） delivered into the subdermal space to treat compromised skin flaps in the rat. Methods Thirty Sprague-Dawley rats weighing 350 -400 g were used in this study. The rats was randomly divided into three groups of ten rats each. A dorsal flap （ 8 cm × 2 cm） in full thickness with the pedicle located at the leve of the iliac crest was designed. Viral transfection with 10^8 plaque-forming units was performed 2 days before the dorsal flap elevation. Rats received subdermal injections of adenovirus encoding VEGF （Ad-VEGF） or green fluorescent protein （Ad-GFP） as treatment control. Another set of animals （n = 10） received PBS was designated as control. Two days after treatment, the flap were elevated as originally designated and sutured back to its bed. The survival rate of the flap was evaluated on day seven after operation. Results The survival rate of the flap in Ad-VEGF group increased significantly as compared with those of the Ad-GFP group（ P 〈 0.01 ）and the PBS group （P 〈 0. 01 ）. Immunohistochemical staining showed that VEGF was expressed in the surviva flap injected with Ad-VEGF. Histological analysis showed that more granulation tissues and angiogenesis were observed in the Ad-VEGF group than those of the Ad-GFP group and the PBS group. Conclusion local application adenovirus-mediated VEGFcDNA may efficiently improve the survival of ischemic skin flap.

关 键 词：内皮生长因子/遗传学 腺病毒科 基因表达 外科皮瓣 

分 类 号：R373.7[医药卫生—病原生物学] R394[医药卫生—基础医学]