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Am J Physiol Regul Integr Comp Physiol 284: R1249-R1254, 2003; doi:10.1152/ajpregu.00490.2002
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Vol. 284, Issue 5, R1249-R1254, May 2003

IL-1beta stimulates IL-6 production in cultured skeletal muscle cells through activation of MAP kinase signaling pathway and NF-kappa B

Guangjo Luo1,*, Dan D. Hershko1,*, Bruce W. Robb1, Curtis J. Wray1, and Per-Olof Hasselgren2

1 Department of Surgery, University of Cincinnati, Cincinnati, and Shriners Hospitals for Children, Cincinnati, Ohio 45267; and 2 Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215

Recent studies suggest that the skeletal muscle may be a significant site of IL-6 production in various conditions, including exercise, inflammation, hypoperfusion, denervation, and local muscle injury. The mediators and molecular mechanisms regulating muscle IL-6 production are poorly understood. We tested the hypothesis that IL-6 production in muscle cells is regulated by IL-1beta and that mitogen-activated protein (MAP) kinase signaling and NF-kappa B activation are involved in IL-1beta -induced IL-6 production. Cultured C2C12 cells, a mouse skeletal muscle cell line, were treated with different concentrations (0.1-2 ng/ml) of IL-1beta in the absence or presence of the p38 MAP kinase inhibitor SB-208350 or the p42/44 inhibitor PD-98059. Protein and gene expression of IL-6 were determined by ELISA and real-time PCR, respectively. NF-kappa B DNA binding activity was determined by electrophoretic mobility shift assay and by transfecting myocytes with a luciferase reporter plasmid containing a promoter construct with multiple repeats of NF-kappa B binding site. Treatment of myotubes with IL-1beta resulted in a dose- and time-dependent increase of IL-6 production accompanied by an ~25-fold increase in IL-6 mRNA levels. IL-1beta stimulated NF-kappa B DNA binding activity and gene activation. SB-208350 and PD-98059 inhibited the increase in IL-6 production induced by IL-1beta . The present results support the concept that skeletal muscle is an important site of IL-6 production. In addition, the results suggest the IL-1beta regulates muscle IL-6 production at least in part by activating the MAP kinase pathway and NF-kappa B.

interleukin-6 production; cytokines; mitogen-activated protein kinase; nuclear factor-kappa B

* G. Luo and D. D. Hershko contributed equally to this work.




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