血管内皮生长因子联合骨髓单个核细胞自体移植治疗大鼠肢体缺血  

Vascular endothelial growth factor combined with bone marrow mononuclear cell autograft for limb ischemia in rats

作　　者：张玉浩[1] 戴坤扬[1] 梅家才[1] 

机构地区：[1]上海交通大学附属第六人民医院外科,上海市200233

出　　处：《中国组织工程研究与临床康复》2007年第15期2882-2885,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:将血管内皮生长因子应用于骨髓单个核细胞自体移植治疗肢体缺血,探索一种简单、安全有效治疗肢体缺血的方法,同时初步分析血管内皮生长因子促进血管新生的机制和疗效。方法:实验于2005-03/2006-07在上海交通大学附属第六人民医院动物实验中心完成。将48只雄性Wistar大鼠采用随机单位组设计分为4组:缺血对照组、血管内皮生长因子治疗组、骨髓单个核细胞治疗组、转染血管内皮生长因子基因的骨髓单个核细胞治疗组,每组12只。结扎所有动物左后肢股动脉及分支,制备后肢缺血模型。以微量加样器自结扎处以下2cm开始,间隔0.2cm为一注射点,共注射于内收肌和腓肠肌5点,每注射点各注射60μL,缺血对照组大鼠注射300μL培养液,血管内皮生长因子治疗组大鼠注射含5μg基因pcDNA3.1-hVEGF165的DNA-脂质体复合物,骨髓单个核细胞治疗组大鼠注射3×106个骨髓单个核细胞,转染血管内皮生长因子基因的骨髓单个核细胞治疗组大鼠注射含3×106个血管内皮生长因子的骨髓单个核细胞。于移植后4周行动脉造影,采用RT-PCR检测血管内皮生长因子基因的体内表达,采用免疫组化法检测缺血区新生血管密度,评价血管新生情况。结果:48只大鼠均造模成功,并进入结果分析。①移植后4周动脉造影显示,缺血对照组大鼠股动脉及其分支均未显影,血管内皮生长因子治疗组和骨髓单个核细胞治疗组可见中等量新生血管,两组效果相似,转染血管内皮生长因子基因的骨髓单个核细胞治疗组可见有大量新生血管新生,丰富的侧枝循环建立。②大鼠毛细血管密度测定结果显示,血管内皮生长因子治疗组和骨髓单个核细胞治疗组均较缺血对照组新生血管数量增加,转染血管内皮生长因子基因的骨髓单个核细胞治疗组可进一步增强疗效[(10.0±0.8),(21.7±1.9),(20.4±3.3),(42.1±3.5)个/HP,P<0.05],血管内皮生AIM： Vascular endothelial growth factor （VEGF） is applied in autologous bone marrow mononuclear cell （BM-MNC） transplantation for limb ischemia. To explore an easier and safer therapy for ischemic arterial disease of the extremity and analyze the mechanism and curative effect of VEGF for accelerating angiogenesis. METHODS： The experiment was conducted in the Animal Experiment Center of Sixth People＇s Hospital of Shanghai Jiao Tong University from March 2005 to July 2006. Totally 48 male Wistar rats were divided randomly into 4 groups： ischemic control group （group A）, VEGF group （group B）, BM-MNC transplantation group （group C）, BM-MNC transfected with VEGF transplantation group （group D）, 12 rats in each group. Left posterior limb artery and branch of all animals ware ligated to establish ischemic models. With micro-pipettor, from 2 cm below ligation point, totally 5 points ware injected on adductor and gastrocnemius with an interval of every 0.2 cm, 60 μL once. 300 μL culture media was injected in the group A. DNA- lipofectamine including 5 μg pcDNA3.1-hVEGF165 Gene was injected in the group B. 3 × 10^8 mononuclear cells ware injected in the group C. Mononuclear cells containing 3 × 10^6 VEGF was injected in the group D. After 4 weeks the expression of VEGF were examined by RT-PCR and the intensity of neovasculorization were evaluated by means of DSA and immunohistochemical method to assess angiogenesis. RESULTS： A total of 48 rats ware made in ischemic model successfully and ware involved in the result analysis. ①After 4 weeks arteriography displayed that in the group A arteriae femoralis and its branch was underdevelopment. In group B and group C middling new vessels could be seen and the two groups had analog quantity. A mass of new vessels and affluent collateral circulation could be seen in group D. ②The capillary density was significantly higher in group B and group C than in group A, and the curative effect was enhanced in the group D [（10.0±0.8）, （21

关 键 词：骨髓干细胞 移植 缺血 细胞因子 

分 类 号：R394.2[医药卫生—医学遗传学]