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Am J Physiol Regul Integr Comp Physiol 279: R1165-R1170, 2000;
0363-6119/00 $5.00
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Vol. 279, Issue 4, R1165-R1170, October 2000

NF-kappa B mediates the protein loss induced by TNF-alpha in differentiated skeletal muscle myotubes

Yi-Ping Li and Micheal B. Reid

Department of Medicine, Baylor College of Medicine, Houston, Texas 77030

Nuclear factor-kappa B (NF-kappa B) regulates the transcription of a variety of genes involved in immune responses, cell growth, and cell death. However, the role of NF-kappa B in muscle biology is poorly understood. We recently reported that tumor necrosis factor-alpha (TNF-alpha ) rapidly activates NF-kappa B in differentiated skeletal muscle myotubes and that TNF-alpha acts directly on the muscle cell to induce protein degradation. In the present study, we ask whether NF-kappa B mediates the protein loss induced by TNF-alpha . We addressed this problem by creating stable, transdominant negative muscle cell lines. C2C12 myoblasts were transfected with viral plasmid constructs that induce overexpression of mutant I-kappa Balpha proteins that are insensitive to degradation via the ubiquitin-proteasome pathway. These mutant proteins selectively inhibit NF-kappa B activation. We found that differentiated myotubes transfected with the empty viral vector (controls) underwent a drop in total protein content and in fast-type myosin heavy-chain content during 72 h of exposure to TNF-alpha . In contrast, total protein and fast-type myosin heavy-chain levels were unaltered by TNF-alpha in the transdominant negative cell lines. TNF-alpha did not induce apoptosis in any cell line, as assessed by DNA ladder and annexin V assays. These data indicate that NF-kappa B is an essential mediator of TNF-alpha -induced catabolism in differentiated muscle cells.

cachexia; cytokine; free radicals; signal transduction; inflammation


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