Objective To illuminate effects of hepcidin on iron metabolism of skeletal muscle. Methods L6 rat skeletal muscle cells were randomly divided into four groups and there were 6 repeats in each group. The cells were incubated with hepcidin at different concentrations (0, 0.05 mg/L, 0.1 mg/L, 0.15 mg/L) for 12 hours. Isotopic tracer method was used to investigate the change of cell iron uptake and iron release. The content of iron in liabile iron pool(LIP) was investigated by flow cytometry and the expression of ferroportin 1(FPN1) in rats’ gastrocnemius was observed by Western blotting. Results Compared with the control group, the released iron of L6 cells which were incubated with 0.1 mg/L hepcidin decreased significantly (P<0.05). There was no significant difference of iron uptake and iron release after incubated with 0.05 and 0.15 mg/L hepcidin (P>0.05). The LIP of L6 cells which were incubated with 0.1 mg/L hepcidin increased significantly (EM>P/EM><0.05). The expression of FPN1 decreased significantly (EM>P/EM><0.05). Conclusion Hepcidin might play an important role in iron metabolism of skeletal muscle cells, which can decrease the expression of FPN1. As a result, the released iron of skeletal muscle cells is reduced and the iron content of ir