电刺激改善下肢缺血及促进骨骼肌血管生成的实验研究

Electrical stimulation relieves ischemia of rat skeletal muscle via angiogenesis

目的 观察电刺激对后肢缺血大鼠VEGF及受体FLK/KDR的调节作用。方法 采用SD大鼠股浅动脉结扎法制备后肢缺血模型 ,应用RT PCR和免疫组化方法检测电刺激对后肢缺血大鼠VEGFmRNA和蛋白表达的影响 ,应用Westernblot和免疫荧光测定FLK 1表达。结果 刺激频率为 2 5Hz ,强度为 0 .3mA ,不引起肌肉收缩。缺血模型建立 7d后连续电刺激 7d ,VEGFmRNA的表达刺激组比非刺激组增加 3倍 (2 .5 8对 0 .93,P <0 .0 1) ,VEGF蛋白增加两倍 (0 .48对 0 .2 4,P <0 .0 1)。FLK 1也有明显增加。结论 电刺激可促进后肢缺血大鼠VEGF和FLK 1/KDR的表达 。

Objective To investigate the effects of electrical stimulation on the expression of vascular endothelial growth factor(VEGF)mRNA and receptor FLK 1/KDR in a ischemic model of rat hindlimb. Methods The model of hindlimb ischemia on the right side was established by ligation of the superficial femoral artery in 10 rats. The rats were then randomized into an experimental group and a control group. The rats in the experimental group were intervened with electrical stimulation ( 25 Hz, 0.1 V) on the tibialis anterior (TA) of the right side, while those in the control group were not. RT PCR and immunohistological methods were used to detect the expressions of VEGF mRNA and protein in TA muscles. FLK 1/KDR was detected by means of Western blot and immunofluorescence. Results After 7 days of continuous stimulation, there was a significant increase in blood flow within the muscle. VEGF mRNA and VEGF protein had 4 fold and 2 fold increases, respectively, in the stimulated TA muscles as compared to the control(2.58 vs 0.93, 0.48 vs 0.24, P <0.01). FLK 1/KDR expression was up regulated in the electrically stimulated skeletal muscle. Conclusion Electrical stimulation could serve as a nonmolecular therapeutic approach for angiogenesis via promoting VEGF and FLK 1/KDR expressions.