腺相关病毒分别介导转化生长因子β_1和血管内皮生长因子联合转染促进糖尿病皮肤溃疡愈合的生物学效应  

Biological effects of co-transfection of TGFβ_1 and VEGF mediated by AAVon promoting healing of the cutaneous ulcer in diabetes mellitus

作　　者：赛佳明 张慧琴[2] 

机构地区：[1]青岛市骨伤医院骨科实验室,山东青岛266021 [2]青岛大学医学院附属医院,山东青岛266003

出　　处：《创伤外科杂志》2007年第5期450-454,共5页Journal of Traumatic Surgery

摘　　要：目的探讨腺相关病毒(AAV)介导转化生长因子β1(TGFβ1)和血管内皮生长因子(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。方法采用四氧嘧啶建立兔糖尿病动物模型,并通过手术方法建立皮肤溃疡创面。将AAV-TGFβ1和AAV-VEGF联合转染糖尿病兔耳皮肤溃疡创面处。通过皮肤溃疡创面愈合大体形态学观测、TGFβ1和VEGF双重免疫组化染色、微循环血管密度显微镜观察计数、主胶原(Ⅰ、Ⅲ型胶原)提取及免疫印迹检测、胶原微量定量检测等方法,研究AAV-TGFβ1和AAV-VEGF联合转基因治疗在促进糖尿病兔耳皮肤溃疡愈合中的生物学效应。结果AAV-TGFβ1和AAV-VEGF联合转基因治疗组与对照组皮肤溃疡创面愈合的检测比较发现:(1)联合转基因治疗组TGFβ1和VEGF基因转录表达增多;(2)联合转基因治疗组创缘毛细血管叉密度(19.18±3.56)个/mm2高于对照组(6.43±1.52)个/mm2,差别有统计学显著性意义(P<0.05);(3)联合转基因治疗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。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并有效地促进溃疡愈合。Objective To observe the biological effects of adeno-associated virus （ AAV） mediated transforming growth factor β1 （AAV-TGFβ1 ） and vascular endothelial growth factor（ AAV-VEGF） on promoting the healing of cutaneous ulcer in diabetic rabbits. Methods The dermal ulcer models of diabetic rabbits were established by injecting alloxan （ 130mg/kgl via ear vein, and the ulcer wound was made. In the co-transfection group, the wound was locally infiltrated,and injected with AAV-TGFβ1 virus and AAV-VEGF virus. The rabbits in the control group were treated with injection of saline. The levels of TGFβ1 and VEGF gene transcription in the healing tissue were detected with polymerase chain reaction（PCR） 1 month postoperatively. The capillary density in the wound margin was counted with microcirculation microscope 3 weeks postoperatively. The collagen Ⅰ and Ⅲ were isolated and detected with western blotting. The content of collagen in the ulcer healing tissue was detected 1 day and 1,2,3 weeks,1,2,3,4,5 and 6 months postoperatively. The thickness,color and quality of the wound healing were observed postoperatively. Results All the 24 rabbits were involved in the analysis of results： （ 1 ） The expressions of TGFβ1 and VEGF gene transcription in the co-transfection group were increased as compared with those in the control group. （ 2） The capillary density at the wound margin in the co-transfecfion group was higher than that in the control group [ （ 19.18 ± 3.56）, （ 6.43 ± 1.52）/mm^2, t = 3.21, P 〈 0.05 ]. （ 3 ） The content of collagen Ⅰ in the ulcer healing tissue in the co-transfection group was obviously increased, and the proportion of collagen I in the constituent ratio of collagen Ⅰ and Ⅲ was increased. The contents of collagen in the ulcer healing tissue at 1 day after transfection were close without significant difference between the two groups （ t = 1.55, P 〉0. 051 ） After that, the contents of collagen in the co-transfection group were all significantly h

关 键 词：糖尿病 溃疡 愈合 转化生长因子 血管内皮生长因子 

分 类 号：R632.103[医药卫生—外科学]