人血管内皮细胞生长因子-165和人血管生成素-1促进大鼠缺血下肢血管新生  

Vascular endothelial grow factor 165 and angiopoitin-1 augment revascularization in a rat hindlimb ischemia model

作　　者：宋建蓉[1] 杨志健[1] 张馥敏[1] 马文珠[1] 哈团柱 李传富 高翔[3] 

机构地区：[1]南京医科大学第一附属医院心脏内科,210029 [2]美国东田纳西州大学James H Quillen医学院外科系 [3]南京大学分子医学研究所

出　　处：《中国介入心脏病学杂志》2005年第2期103-107,共5页Chinese Journal of Interventional Cardiology

基　　金：江苏省科委自然科学基金(BK2001118);江苏省卫生重点项目(H2004)

摘　　要：目的　观察肌肉转染腺病毒(adenovirus ,Ad )携带的人血管内皮细胞生长因子- 1 6 5(ves cularendothelialgrowfactor ,Ad5 VEGF1 65)和人血管形成素- 1 (angiopoietin- 1 ,Ad5 Ang -1 )基因对大鼠缺血下肢的生血管效应。方法　制作大鼠下肢血管闭塞模型,肌肉内注射Ad5 VEGF1 65 或 和Ad5 Ang -1 ,以Westernblot法检测生长因子蛋白表达,免疫组化检测基因导入后在缺血肌肉引起的效应。结果　肌肉经注射Ad5 VEGF1 65和Ad5 Ang- 1后高表达人VEGF1 65 蛋白和人Ang- 1蛋白。术后7d处理组与对照组新生毛细血管数目无明显差别。但术后1 4d和2 1dAng- 1和VEGF1 65 组的新生毛细血管 肌肉数目比明显高于对照组(P <0 . 0 1 ) ,VEGF1 65+Ang 1两因子合用组明显高于单用组(P <0. 0 1 )。Ad -VEGF1 65组及Ad VEGF1 65+Ad Ang 1合用组出现大量包围着一厚层αSMA阳性平滑肌细胞的血管,其数量与肌肉数比值明显高于对照组(P <0 . 0 1 )。因子单用组和合用组肌肉内出现大量溴化脱氧尿嘧啶阳性细胞浸润,部分细胞表面呈现C- Kit阳性,新生血管内皮细胞中也有C Kit阳性标记。结论　Ad5- VEGF1 65和Ad5 -Ang -1能促进缺血肢体血管新生,二者合用的生血管效应更为显著;VEGF1Objective To study the angiogenic effects of adenovirus mediated human vascular endothelial growth factor 165 and human angiopoietin-1 (Ad 5-VEGF 165 , Ad 5-Ang-1) in rat models of hindlimb ischemia. Methods Rat models of hindlimb ischemia were established by ligation and peeling off rat′s femoral arteries. Ad-VEGF 165 and Ad-Ang-1 were intramuscularly transferred into the rat ischemic hindlimbs. The expression of VEGF 165 and Ang-1 were examined by Western blotting. Immunohistochemistry was performed to illustrate the effects on rat ischemic muscles after gene transferring. Results (1) Western blotting showed a high expression of VEGF 165 and Ang-1 in the ischemic hindlimb muscle transferred with Ad 5-VEGF 165 and Ad 5-Ang-1 VEGF 165 respectively. (2) There was no significant difference between groups on 7 days after the transfer. The capillary to muscle fiber ratio was significantly higher in the treating groups than that in control groups and were significantly higher in VEGF 165 +Ang-1 group than in VEGF 165 or in Ang-1 groups on day 14th and day 21th. (3) Many structured vessels surrounded by a layer of smooth muscle cells were found in Ad-VEGF 165 and Ad-VEGF 165 +Ad-Ang-1 groups, the number of SMA + vessel per muscle fiber was obviously higher than those in control groups. (4) Enormous cells positive for BrdU appeared in treated muscles in Ang-1, VEGF 165 , VEGF 165 +Ang-1 groups and many of them were positive for C-Kit, an antigen expressed by pluripotent marrow stem cells. Some C-Kit+ cells were incorporated in sites of neovascularization. Conclusion (1) Vascular endothelial growth factor 165 and angiopoietin-1 can promote neovascularization in rat models of hindlimb ischemia and the angiogenic effect is much more significant in Ad-VEGF+Ad-Ang-1 group. (2) VEGF 165 can increase the number of vessles that are coated with smooth muscle cells which shapes are similar to arterioles. (3) Not only angiogenic effect but perhaps vasculogenic effct also

关 键 词：血管内皮细胞生长因子 血管新生 人血管生成素-1 缺血下肢 大鼠 Ad-VEGF165 人VEGF165 溴化脱氧尿嘧啶 Ang-1 平滑肌细胞 C-Kit factor 下肢血管闭塞 WESTEM 免疫组化检测 新生毛细血管 阳性细胞浸润 血管生成作用 血管效应 

分 类 号：R735.7[医药卫生—肿瘤] R541.4[医药卫生—临床医学]