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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-165(vascularendothelialgrowfactor,Ad-VEGF165)和人血管形成素-1(angiopoietin-1,Ad-Ang-1)基因对大鼠缺血下肢的生血管效应。方法:用SD雄性大鼠84只,分为7个组,每组12只,分别为正常组、假手术组、结扎组、绿色荧光蛋白(greenfluorescenceprotein,GFP)组(作为病毒对照组)、Ang1组、VEGF165组、Ang1+VEGF165组。制作大鼠下肢血管闭塞模型,肌肉内注射Ad-VEGF165或/和Ad-Ang-1,以Western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:①肌肉经注射Ad-VEGF165和Ad-Ang1后高表达人VEGF165蛋白和人Ang1蛋白。②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14d和21dAng1和VEGF165组的毛细血管/骨骼肌纤维数目比明显高于对照组(F=24.53,P<0.01),VEGF165+Ang1两因子合用组明显高于单用组(F=12.22,P<0.01)。③VEGF165组及VEGF165+Ang1合用组出现大量包围着一厚层平滑肌肌动蛋白α(smoothmuscleactin,α-SMA)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(F=9.73,P<0.01)。④因子单用组和合用组肌肉内出现大量BrdU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
OBJECTIVE: To observe the expression of Ad-VEGF165 and Ad-Ang-1 gene in adenovirus (Ad-) Vascularization effect on ischemic limbs in rats. Methods: Eighty-four male Sprague Dawley rats were randomly divided into seven groups (n = 12 each): normal group, sham operation group, ligation group, green fluorescent protein (GFP) Group, VEGF165 group, Ang1 + VEGF165 group. The model of lower extremity vascular occlusion was made. Ad-VEGF165 and / or Ad-Ang-1 were injected intramuscularly. The expression of growth factor protein was detected by Western blot. The effect induced by ischemic muscle was detected by immunohistochemistry. RESULTS: Muscle injected with Ad-VEGF165 and Ad-Ang1 highly expressed human VEGF165 protein and human Ang1 protein. ② There was no significant difference in the number of newborn capillaries between the treatment group and the control group 7 days after operation. However, the number of capillary / skeletal fibers in Ang1 and VEGF165 group at 14d and 21d after operation was significantly higher than that in control group (F = 24.53, P <0.01), VEGF165 + P <0.01). (3) There was a large number of blood vessels surrounding the smooth muscle actin (α-SMA) -positive smooth muscle cells in the VEGF165 group and the VEGF165 + Ang1 group, with a significantly higher ratio of the number to the number of muscles (F = 9.73, P <0.01). (4) A large number of BrdU-positive cells infiltrated in muscle of single-use and combined-use groups, C-Kit was positive on some cells and C-Kit