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Abstrac: Continuous low-voltage electrical stimulation (CL-VES) of skeletal muscle at a level far below the threshold of muscle contraction has been reported to induce synthesis of vascular endothelial growth factor (VEGF) protein and promote local angiogenesis in a hindlimb ischemia model of rats. However, the mechanism of the induction of VEGF protein synthesis and promotion of local angiogenesis in skeletal muscle by CL-YES has not been fully understood. Electrical stimulation (ES) by electrodes in skeletal muscle may cause hypoxia and muscle damage. Therefore, in the present study, we examined whether the CL-YES-induced VEGF production in skeletal muscle in hindlimb ischemia model of rats was due to hypoxia or inflammation by immunohistorical method with anti-hypoxiainducible factor a (HIFa) antibody and interleukin 6 (IL 6) antibody in tissues. HGF is more potent than VEGF in the role of collateral formation and has an anti-apoptosis function. We also investigated whether other angiogenic factors, such as hepatocyte growth factor (HGF) and fibroblast growth factor (FGF) that induced VEGF also contribute to the local angiogenesis by LVES. We also elucidated the mechanism of the induction of local angiogenesis in skeletal muscle of hindlimb ischemia model of rats by ES with special reference to the interaction of strength, duration of ES and VEGF and hepatocyte growth factor (HGF). HGF induces VEGF. Not only VEGF but also HGF was significantly increased in tissues by LVES. There was no difference in the level of immunohistochemical staining of the tibialis anterior (TA) muscles with polyclonal anti FGF antibody, anti IL 6 antibody, and HIF antibody between the CL-YES group and control group. Although CL-YES induced VEGF earlier than IH-YES, the VEGF level of all 3 ES groups increased to a similar level at 5 days, suggesting that whichever 3 electrical stimulation were useful to induce VEGF. However, the levels of HGF were statistically almost the same between stimulated muscle and control in the IH-YES groups. These results suggest that CL-YES in skeletal muscle was effective for VEGF induction and HGF induction which was not seen with IH-YES and that the CL-YES-induced VEGF production in this model might not be due to hypoxia or inflammation by CL-YES. Therefore CL-YES might be a useful technique in clinical applications for ischemic vessel deseases such as arteriosclerosis obliterance. Moreover, there is a possibility that the mechanisms of angiogenesis by CL-YES and IH-YES may be different (Jpn J Rehabil Med 2002; 39: 457-466)

Angiogenic Growth Factors in Rat Ischemic Skeletal Muscle by Continuous Low-voltage Electrical Stimulation.