转染血管内皮生长因子基因的骨髓间充质干细胞修复糖尿病大鼠足背创面

背景：糖尿病足溃疡严重威胁患者的健康甚至生命，其治疗仍是国际性难题，目前缺乏良好有效的治疗手段，而基因疗法和干细胞疗法在创伤修复领域展现出独特的优势和潜力。目的：探索转染人血管内皮生长因子165（human vascular endothelial growth factor 165, hVEGF165）基因的骨髓间充质干细胞对大鼠糖尿病足背创面的治疗效果。方法：体外构建hVEGF165基因的腺病毒表达载体，转染第3代大鼠骨髓间充质干细胞；将120只雄性Wistar大鼠分成正常溃疡对照组、模型组、基因治疗组、干细胞治疗组和基因转染干细胞治疗组。后4组大鼠经腹腔注射链脲佐菌素诱导糖尿病，在所有大鼠的后足背切取一3 mm×7 mm矩形全层皮肤制成足背创面模型。各组大鼠均在足背创缘皮下分点注射移植，距创缘5 mm处、等距6点，正常溃疡对照组和模型组各点均注射50μL的PBS，基因治疗组各点注射50μL的腺病毒悬液（1×1013 pfu/L），干细胞治疗组各点注射50μL的干细胞悬液（1×1010 L-1），基因转染干细胞治疗组各点注射50μL的病毒转染后的干细胞悬液（1×1010 L-1）。结果与结论：与模型组、基因治疗组、干细胞治疗组相比，基因转染干细胞治疗组大鼠创面愈合率、血管内皮生长因子mRNA及蛋白表达水平、局部毛细血管数量均有显著提高（P〈0.05），但较正常溃疡对照组差（P〈0.05）。说明 hVEGF165基因联合骨髓间充质干细胞疗法能够促进大鼠糖尿病足背创面的愈合，其主要机制为促进局部血管的生成及血管内皮生长因子的表达。

Transplantation of bone marrow mesenchymal stem cells transfected by vascular endothelial growth factor gene promotes foot wound healing in diabetic rats

BACKGROUND：Diabetic foot ulcers threaten the patients’ health and even survival seriously. It is an international difficult problem and lacks an effective treatment. But gene therapy and stem celltherapy possess special advantages and potential in wound healing. OBJECTIVE：To assess the therapeutic effect of transplantation of bone marrow mesenchymal stem cells transfected by human vascular endothelial growth factor 165 （hVEGF165） gene on foot wound healing in diabetic rats. METHODS：Recombinant adenovirus was established in vitro which expressed hVEGF165 gene and transfected into the third generation of bone marrow mesenchymal stem cells. Total y 120 male Wistar rats were divided into five groups：group A （non-diabetic controls）, group B （diabetic controls）, group C （Ad-hVEGF165 therapy）, group D （stem celltherapy） and group E （transplantation of bone marrow mesenchymal stem cells transfected by Ad-hVEGF165 gene）. Rats in the latter four groups were intraperitoneal y injected with streptozotocin to induce diabetic models. In al rats, a 3 mm×7 mm rectangular ful-thickness skin sample was cut from the instep of the hind foot to make a model of foot wound. The rats were subcutaneously injected at equidistant six points 5 mm＆amp;nbsp;distal to the wound edge on the dorsum of the foot：50μL PBS per point for group A, 50μL adenovirus suspension （1×1013 pfu/L） per point for group C, 50μL stem cellsuspension （1×1010/L） per point for group D, and 50μL adenovirus suspension＋50μL stem cellsuspension per point for group E. RESULTS AND CONCLUSION：After injection, the rate of wound healing, the expression of VEGF and the qualities of capil aries in group E were higher when compared with groups B, C, D （P〈0.05）, but were lower than those in group A （P〈0.05）. Transplantation of bone marrow mesenchymal stem cells transfected by hVEGF165 gene can promote foot wound healing, angiogenesis and expression of VEGF in diabetic rats.