腺病毒介导血管内皮生长因子基因促进大鼠随意皮瓣的早期断蒂  

Adenovirus-mediated vascular endothelial growth factor gene in promotion of early pedicle division of random flap in rats

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

机构地区：[1]安徽医科大学第一附属医院整形外科,安徽省合肥市230022

出　　处：《中国临床康复》2006年第21期63-65,共3页Chinese Journal of Clinical Rehabilitation

基　　金：安徽省科技厅自然科学基金资助项目(03023056)~~

摘　　要：目的:探讨腺病毒介导血管内皮细胞生长因子165基因转染大鼠皮瓣,对皮瓣早期断蒂的影响。方法:实验于2004-08/2005-09在安徽医科大学第一附属医院外科动物实验中心及中心实验室完成。健康SD雄性大鼠30只,随机分成3组:目的组、绿色荧光蛋白对照组、空白对照组,每组10只。所有实验大鼠背部设计2cm×6cm随意皮瓣,蒂位于髂脊水平。皮瓣形成后,术时即刻将磷酸盐缓冲液稀释的重组复制缺陷型腺病毒携带血管内皮细胞生长因子165基因溶液(目的组)、磷酸盐缓冲液稀释的重组复制缺陷型腺病毒携带绿色荧光蛋白溶液(荧光蛋白对照组)、磷酸盐缓冲液溶液(空白对照组)1mL注射到大鼠皮瓣内,给药后即刻以3-0丝线原位缝合,继续单笼饲养。所有实验大鼠皮瓣4d后断蒂,断蒂后7d观测皮瓣存活面积,取皮瓣远端存活区组织苏木精-伊红染色,光镜下观察组织学变化,免疫组织化学染色检测血管内皮细胞生长因子165表达情况,用CD34免疫组织化学染色计算血管密度。结果:①断蒂后7d,所有实验动物全部存活,皮瓣存活区与坏死区已经明确。②皮瓣存活率、皮瓣血管断面密度目的组和对照组之间差异有显著性犤皮瓣存活率:目的组为(98.8±1.5)%,荧光蛋白对照组为(78.3±2.5)%,空白对照组为(79.2%±2.4)%;皮瓣血管断面密度:目的组为(49.6±6.3)个/高倍视野,荧光蛋白对照组为(37.5±3.0)个/高倍视野,空白对照组为(39.5±3.0)个/高倍视野,P<0.05犦,对照组之间差异没有显著性(P>0.05)。③免疫组织化学染色各组均可见棕黄色颗粒沉积,目的组比对照组染色深。④组织学观察,目的组皮瓣真皮、肉芽及肌肉中可见大量毛细血管,而对照组较少。结论:应用重组复制缺陷型腺病毒携带血管内皮细胞生长因子165基因转染大鼠皮瓣具有促进皮瓣早期断蒂的作用。AIM： To investigate the effect of adenovirus vascular endothelial growth factor 165 （Ad-VEGF165） gene transfection on early pedicle division of random flap in rats. METHODS： The experiment was conducted in the Surgical Experimental Animal Center and Central Experimental Laboratory of First Hospital Affiliated to Anhui Medical University from August 2004 to September 2005. Thirty male healthy Sprague-Dawley rats were randomly divided into three groups： destination group, green fluorescent protein group （GFP group） and blank control group with 10 rats in each gr6up. A 2 cm×6 cm dorsal random skin flap with pedicle at the iliac crest was designed on all of the experimental rats. After the skin flap formed, 1 mL of replication-deficient recombinant Ad-VEGF165 diluted by phosphate buffer （PB）, replicationdeficient recombinant adenovirus GFP （Ad-GFP） diluted by PB and PB were respectively injected into the skin flap of rats in the destination group, GFP group and blank control group immediately at the operation. As soon as administration, a 3-0 suture silk was used to conduct suturation in situ, and rats was then raised separately. The pedicles of all rats were divided 4 days later. On the 7^th day after pedicle division, the survival rate of all skin flaps were investigated and skin samples in the distal survival portion of flaps were stained with hematoxylin and eosin, whose histological changes were observed under light microscope. The expression of VEGF165 was detected with immunohistochemical stain, and the density of capillary was calculated by CD34 immunohistochemical stain. RESULTS： ①At 7 days after pedicle division, all the experimental animals survived, and the survival and necrotic zones were identified.②Differences in survival rate and density of capillary sections in flaps were remarkable between the destination group and control group [Survival rates of destination group, GFP group and blank control group were （98.8 ±1.5）%, （78.3±2.5）% and （79.2%±2.4）% respe

关 键 词：腺病毒科 血管 内皮生长因子 基因疗法 外科皮瓣 

分 类 号：R318[医药卫生—生物医学工程]