复制缺陷型腺病毒载体介导的VEGF基因治疗大鼠缺血皮瓣的研究

The effect of adenover-mediated VEGF gene on surival of a random skin flap in the rat

目的探讨重组复制缺陷型腺病毒携带VEGF基因对大鼠缺血皮瓣成活的影响。方法选用30只SD大鼠,体重350～400 g,随机分为三组,Ad-VEGF组、Ad-GFP组和空白对照组,每组10只,在其背部形成8 cm×2 cm的全厚随意型皮瓣。于皮瓣受区筋膜下分别注射携带血管内皮细胞生长因子基因的腺病毒(Ad-VEGF_(165)cDNA)、携带绿色荧光蛋白基因的腺病毒(Ad-GFP)、0.9%氯化钠注射液。皮瓣原位缝合,术后7 d观察实验结果。结果基因组皮瓣成活面积百分比与其他两个对照组差异有统计学意义。免疫组织化学显示VEGF_(165)组毛细血管周围VEGF沉积、组织切片微血管密度明显高于两个对照组(P<0.05)。结论局部受区筋膜下注射Ad-VEGF_(165)可诱导局部VEGF蛋白表达,促进新生血管的形成,提高缺血皮瓣的成活面积,是一种简单有效的基因治疗方法。

Objective To investigate the effect of adenovirus-medicated gene therpy with vascular endothelial growth factor（VEGF） delivered into the subdermal space to treat compromised skin flap in the rat. Methods Thirty sprague-dawley rats weighing 350 - 400g were used in this study. The rats was randomly divided into three groups of ten rats each. A dorsal flap（8cm × 2cm） in full thickness with the pedicale located at the leve of the illaccrest was designed,each,animal received recipient bed subfascial injections of adenovirus encoding VEGF（Ad-VEGF） or green fluorescent protein（Ad-GFP） as treatment control. Another set of animals（ n = 10） received NG was designated as control. The flap were elevated as originally designated and sutured back to its bed. The survival rate of the flap was evaluated on day seven after operation. Results The survival rates of the flap in Ad-VEGF group increased significantly as compared with those of the Ad-GFP group（ P〈 0.05 ） and the NG group（ P 〈 0.005 ）. Immunohistochemical staining showed that VEGF was expressed in the surviva flap injected with Ad-VEGF. Histological analysis showed that more granulation tissue and angiogenesis were observed in the Ad-VEGF group than those of the Ad-GFP group and the NG group. Conclusion The results may indicate that advector carrying VEGFcDNA could be useful in enhancing the survival of the skin flap due to the effect of the local delivery of the VEGF.