Ad-EGFP修饰MSCs在糖尿病足溃疡大鼠体内示踪及疗效观察.docVIP

Ad-EGFP修饰MSCs在糖尿病足溃疡大鼠体内示踪及疗效观察[摘要]目的：观察自尾静脉移植Ad-EGPF修饰的大鼠骨髓间充质干细胞(BMSCs)在糖尿病足溃疡模型大鼠胰腺、肾脏及溃疡局部的示踪情况，并观察移植后大鼠血糖、体重及溃疡面积的改变情况。方法：贴壁法体外分离培养Wistar大鼠BMSCs，传代培养后用携带增强型绿色荧光蛋白基因的腺病毒(Ad-EGFP)以感染复数(MOI=150)体外转染。60 只Wistar大鼠，雌雄各半，随机分为3 组；糖尿病治疗组(糖尿病足溃疡大鼠细胞移植治疗组)、糖尿病对照组(糖尿病足溃疡大鼠PBS对照组)使用链脲菌素(STZ)诱导糖尿病模型后，连同正常对照组(正常足溃疡大鼠PBS对照组)制作足部溃疡模型。造模成功后，糖尿病治疗组经尾静脉注射转染Ad-EGFP 72h后的BMSCs悬液0.5 ml(细胞数5×106个)，糖尿病对照组及正常对照组注射等量PBS溶液。分别于细胞移植后7 天、14 天、21 天测量体重、空腹血糖水平、足部溃疡面积；并于细胞移植后14 天，观察BMSCs在胰腺、肾脏及足溃疡局部的分布情况。结果：移植后14天可在糖尿病大鼠胰腺、肾脏及皮肤溃疡面观察到Ad-EGFP标记的BMSCs. 糖尿病治疗组大鼠体重明显恢复、血糖显著降低、足溃疡愈合加速，与糖尿病对照组相比，差异均具有统计学意义(P＜0.05)。结论：BMSCs具有归巢特性，并可能对糖尿病及足溃疡有一定的治疗效果。 [关键词]骨髓间充质干细胞；糖尿病；溃疡；移植；归巢 [中图分类号]R332[文献标识码]A[文章编号]1008-6455（2011）10-1571-04 Study of diabetic foot ulcer transplanted with bone marrow mesenchymal stem cells transfected with Ad-EGFP in rats LIANG Wen-Jia1, CAI Qian2, LIU Yi2 (1.Second Clinical Medical College, Lanzhou University,Lanzhou 730000,Gansu,China; 2.Department of Burns and Plastic Surgery, Lanzhou General Hospital of Lanzhou Area Command of Chinese PLA,Lanzhou 730050,Gansu,China) Abstract:ObjectiveTo investigate the distribution of rat bone marrow mesenchymal stem cells (BMSCs) transfected with Ad-EGFP in kidney and pancreatic tissues, and the variation of blood glucose, weight and ulcer area after cell transplantation, in model rats of diabetic foot ulcer.MethodsMSCs isolated from Wistar rats were cultured in vitro and were then transfected with green fluorescent protein (Ad-EGFP) at the multiplicities of infection (MOI) of being 150. The BMSCs were obtained for transplantation. Sixty Wistar rats were randomly assigned to 3 groups: diabetic treatment group (diabetic foot ulcer rat treated with the transfected BMSCs), diabetic control group (diabetic foot ulcer rat treated with PBS) and normal control group (normal ulcer rat treated with PBS). The two groups of diabetic rat model were achieved by intraperitoneally injection of streptozotocin. Foot ulcer for the three groups w