hVEGF基因工程生物膜对全层皮肤缺损创面中flt-1和TSP-1表达的影响

Effects of hVEGF genetically engineering membrane on the expression of flt-1 and TSP-1 during the healing process of full-thickness cutaneous defect

目的探讨人血管内皮细胞生长因子(hVEGF)基因工程生物膜对创面组织fms样酪氨酸酶(flt-1)和血小板反应蛋白-1(TSP-1)表达的影响。方法建立hVEGF基因工程生物膜覆盖大鼠全层皮肤缺损创面的动物模型,将80只SD大鼠随机分为A、B、C、D四组,每组20只,所有动物于背部制备皮肤全层缺损创面。A组:创面覆盖hVEGF基因工程生物膜和Tegaderm切口膜;B组:创面覆盖空白生物膜和Tegaderm切口膜,C组:创面覆盖含空质粒的成纤维细胞种植的生物膜和Tegaderm切口膜;D组:创面覆盖Tegaderm切口膜。采用免疫组化和图像分析的方法,观察各组在术后3、7、14、29d时创面中flt-1和TSP-1表达的情况。结果术后3、7、14d时,A组flt-1阳性细胞数显著高于B、C、D组(P<0.05或P<0.01);术后7、14、29d时A组创面中TSP-1表达显著低于B、C、D组(P<0.01)。结论将hVEGF基因工程生物膜覆盖于大鼠全层皮肤缺损创面,能使创面持续表达flt-1并抑制TSP-1表达,从而促进新生血管形成,有利于创面愈合。

Objective To study the effects of hVEGF on the expression of fms-like tyrosine kinase-1 （fit-1） and thrombospondin 1 （TSP-1） during the healing process of rats＇ full-thickness cutaneous defect which covered by genetically membrane planted by fibroblast integrated with hVEGF recombinant. Methods Wounds with full-thickness cutaneous defect were produced on the dorsum of the SD rats. Eighty rats were divided into four groups at random, 20 rats each. For the rats in the experimented group A, the wounds were covered with hVEGF genetically engineering membrane and Tegaderm membrane. In the control group B, the wounds were covered with membrane planted by the fibroblasts integrated with blank plasmid and Tegaderm membrane. In the control group C, the wounds were covered with blank membrane and Tegaderm membrane. In control group D, the wounds were only covered with Tegadern membrane. Specimens were obtained at the 3,7,14 and 29 days after injury, immunohistochemistry and image analysis were used to observe the expression of fit-1 and TSP-1. Results The expression of fit-1 in vascular endothelial cells was increased obviously in the experimental group. There was significant difference between the experimental group and other groups at 3, 7 and 14d after injury （ P 〈 0.05 or P 〈 0.01 ）. The mean optical density value was used to reveal the expression level of TSP-1. At the 7, 14 and 29 days after injury, the expression level of TSP-1 was significant lower in experimental group than in control groups （ P 〈 0. 01 ）. Conclusion The application of the hVEGF genetically engineering membrane on the full-thickness cutaneous defect increase the expression of fit-1 and inhibit the expression of TSP-1. This could increase angiogenesis and accelerate the healing process.