腺相关病毒介导转化生长因子β1和血管内皮生长因子联合转染促进糖尿病溃疡愈合的生物学效应(英文)  

Biological effects of co-transfection of transforming growth factor beta 1 and vascular endothelial growth factor mediated by adeno-associated virus on promoting the dermal ulcer healing in diabetes mellitus

作　　者：赛佳明[1] 张慧琴[2] 

机构地区：[1]青岛大学医学院,山东省青岛市266003 [2]青岛大学医学院附属医院妇科,山东省青岛市266003

出　　处：《中国临床康复》2006年第48期213-215,F0003,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：背景:糖尿病患者溃疡创面难愈合,目前认为是由糖尿病患者创面微循环障碍以及内源性生长因子含量降低等因素所造成。目的:观察腺相关病毒介导转化生长因子β1(AAV-TGFβ1)和血管内皮生长因子(AAV-VEGF)联合转基因治疗促进糖尿病皮肤溃疡愈合的生物学效应。设计:随机对照动物实验。单位:青岛大学医学院和青岛大学医学院附属医院。材料:实验于2004-07/2006-01在青岛大学医学院附属医院妇科实验室进行。选用成年健康大白兔24只,单纯随机分为联合转基因组和对照组各12只。方法:①采用耳缘静脉注入四氧嘧啶(130mg/kg)建立糖尿病模型,并通过手术方法建立溃疡创面。②联合转基因组创面局部浸润注射AAV-TGFβ1和AAV-VEGF(9×106/mL病毒颗粒);对照组注射生理盐水。主要观察指标:①术后1个月,通过聚合酶链反应检测愈合组织中转化生长因子β1和血管内皮生长因子基因的转录水平。②术后3周,用微循环显微镜计数创缘毛细血管密度值。③术后6个月通过蛋白凝胶电泳和半干电转移法对Ⅰ、Ⅲ型胶原进行分离及免疫印迹检测。④术后1d,1,2,3周,1,2,3,4,5,6个月时测定溃疡愈合组织中胶原含量。⑤术后观测创面愈合厚度、颜色、质地。结果:24只兔全部进入结果分析。①联合转基因组转化生长因子β1和血管内皮生长因子基因转录表达增多。②联合转基因组创缘毛细血管密度值高于对照组[(19.18±3.56),(6.43±1.52)个/mm2,P<0.05]。③联合转基因组愈合组织中胶原含量均增多(P<0.05),而且Ⅰ型和Ⅲ型胶原构成比中Ⅰ型胶原的比例增加。④在转染后1d,联合转基因组与对照组溃疡愈合组织中的胶原含量接近(P>0.05)。此后,联合转基因组胶原含量均高于对照组(P<0.05)。⑤对照组溃疡愈合明显滞后,愈合质量差。结论:AAV-TGFβ1和AAV-VEGF可联合高效转染糖尿病兔耳皮肤溃疡创面,可使溃疡组织中�BACKGROUND： The ulcer wound is hard to heal in diabetic patients, and it is believed to be caused by the microcirculatory disorder of wound and decreased contents of endogenous growth factors in patients with diabetes mellitus. OBJECTIVE： To observe the biological effects of adeno-associated virus （AAV） mediated transforming growth factor beta1 （AAV-TGFβ1） and vascufar endothelial growth factor （AAV-VEGF） in promoting the dermal ulcer healing of diabetic rabbits. DESIGN： A randomized controlled animal experiment. SETTINGS： Medical CoUege, Qingdao University; Affiliated Hospital of Medical College, Qingdao University. MATERIALS： The experiments were carried out in the gynecological laboratory, Affiliated Hospital of Medical College, Qingdao University from July 2004 to January 2006. Twenty-four healthy adult New Zealand rabbits were randomly divided into co-transfection group （n=12） and control group （n=12）. METHODS： ① The dermal ulcer models of diabetic rabbits was established by injecting alloxan （130 mg/kg） via ear vein, and the ulcer wound was made by operation.② In the co-transfection group, the wound was locally infiltrated, and injected with AAV-TGFβ1, virus and AAV-VEGF virus （the concentration was 9×10^6 virus granules/mL respectively）. The rabbits in the control group were treated with injection of saline. MAIN OUTCOME MEASURES： ① The levels of TGFβ1, and VEGF gene transcription in the healing tissue were detected with polymerase chain reaction （PCR） at 1 month postoperatively. ② The capillary density in the wound margin was counted with microcireulation microscope at 3 weeks postoperatively. ③ The collagenⅠ and Ⅲ were isolated and detected with Western blotting by protein gel electrophoresis and semi-dry electrophoretic transfer. ④ The content of collagen in the ulcer healing tissue was detected at 1 day, 1, 2 and 3 weeks, 1, 2, 3, 4, 5 and 6 months postoperatively.⑤ The thickness, color and quality of the wound healing were observed po

关 键 词：糖尿病/并发症 伤口愈合 转染 转化生长因子β 血管内皮生长因子类 

分 类 号：R587.1[医药卫生—内分泌]