超声微泡介导hVEGF165基因靶向转染糖尿病鼠缺血骨骼肌

Ultrasound-mediated Microbubbles Destruction Enhanced h VEGF165 Gene Expression in Ischemic Skeletal Muscle of Diabetic Rats

目的:探讨超声介导微泡破裂法促进血管内皮生长因子(VEGF)基因在糖尿病鼠缺血骨骼肌内转染的作用,评估其转染效率和安全性。方法:建立糖尿病鼠缺血骨骼肌动物模型,以绿色荧光蛋白基因为报告基因,观察接受超声及微泡治疗组h VEGF165基因在糖尿病鼠缺血骨骼肌内表达,并与对照组相比。同时取糖尿病鼠缺血骨骼肌进行HE染色行组织学检查。结果:在超声介导微泡破裂组内,h VEGF165基因表达明显增强(42.87±5.12),与单纯接受质粒治疗组(5.02±1.21)和接受质粒和超声治疗组(8.16±2.43)相比,差异具有统计学意义(P<0.001),HE切片未发现肌组织结构的改变。结论:超声介导微泡破裂法能有效促进外源基因在糖尿病鼠缺血骨骼肌中表达,为糖尿病周围血管疾病的基因治疗提供了实验依据。

Objective： To evaluate the efficiency and safety of the method of ultrasound-mediated microbubbles destruction to enhance vascular endothelial growth factor（VEGF） gene transfecting in ischemic skeletal muscle of diabetic rat. Methods： To establish an animal model of ischemic skeletal muscle in diabetic rat, green fluorescent protein as reporter gene, observe h VEGF165 gene expression in ischemic skeletal muscle in diabetic mice in different group, and histological study of skeletal muscle was performed after experiments to evaluate the potential effects of gene transfer. Results： In the ultrasound-mediated microbubbles destruction group,h VEGF165 gene expression was significantly increased（42.87±5.12） and compared with the other two groups（5.02±1.21, 8.16±2.43,respective）, the difference was statistically significant（P〈0.001）, whereas the histological analyses did not reveal harmful effect.Conclusion： The method of ultrasound-mediated microbubbles destruction can effectively promote h VEGF165 gene expression in ischemic skeletal muscle in diabetic mice, and this method provides an experimental basis for gene therapy of diabetic peripheral vascular disease.