腺病毒载体介导人VEGF cDNA促进随意型皮瓣成活的实验研究

Effects of hVEGF cDNA on random skin flap via a replication-deficient adenovirus vector

目的探讨以腺病毒载体局部应用血管内皮细胞生长因子基因对大鼠缺血随意型皮瓣成活的影响。方法30只SD大鼠随机分成3组,每组10只,于鼠背设计蒂在尾侧髂嵴水平的随意型皮瓣,面积为2cm×8cm,A组皮瓣远端23顺皮瓣长轴分10个点真皮下注入1012PFU的携带人血管内皮细胞生长因子基因的腺病毒(AdCMVVEGF),B组同法注入同剂量的携带β半乳糖苷酶基因的腺病毒(AdCMVGal),C组注入1ml的生理盐水,3d后掀起皮瓣并原位缝合,术后7d观察实验结果。结果每组皮瓣成活面积的百分比分别为A组(8591±254)%,B组为(5956±118)%,C组为(6148±109)%,A组成活率显著高于B组和C组,差异有非常显著性意义(P<001),B组与C组比较差异无显著性意义(P>005)。原位杂交、免疫组化显示,应用AdCMVVEGF治疗的成活皮瓣中,VEGF有表达,组织学切片显示,AdCMVVEGF治疗皮瓣中肉芽组织增生,新生血管大量形成。结论以腺病毒为载体局部应用VEGF基因能增加缺血皮瓣的成活面积,该法具有高效的特点。

Objective\ The purpose of this study was to determine the effects of local delivery of vascular endothelial growth factor(VEGF)transferred with adenovirus\|mediated gene on the survival of ischemic random skin flap in rats. Methods\ The animals were divided into three groups randomly( n =10).A 2 cm×8 cm dorsal skin flap was designed with the pedicle at the level of the iliac crest.In group A (AdCMV\|VEGF),each animal received 10 12 PFU replication\|deficient recombinant adenovirus(AdCMV\|VEGF)in the distal two\|thirds of the proposed flap by means of the subdermal injection at ten different locations.In group B (AdCMV\|Gal),each received 10 12 PFU AdCMV\|Gal.In Group C (Saline),each received 1 ml saline.Three days after the treatment,the flap was elevated as planed way and re\|sutured back to its donor site.All the animals were evaluated 7 days after the operation. Results\ The mean percentage of surviving flap area was(85.91±2.54)% in group A,(59.56±1 18)% in group B,and (61 48±1 09)% in group C.There was a significant increase in the percentage of the survival area in the flaps of the group A,compared with the group B and group C(Group B vs.Group A, P <0 01;Group C vs.Group A, P <0 01,Group B vs.Group C, P >05).Hybridization in the situ,the immunohistochemical stain showed that the VEGF was expressed in the survival tissue of the flap treated with the AdCMV\|VEGF,but it was not found in the control groups.Histological analysis demonstrated qualitatively greater amount of granulation tissue and angiogenesis was found in the group treated with the AdCMV\|VEGF than the controls. Conclusions\ The results may indicate that Ad vector carrying VEGF cDNA could be useful in enhancing the survival of the skin flap due to the effect of the local delivery of the VEGF.\;