Tumor necrosis factor-α inhibits myogenesis through redox-dependent and -independent pathways

Abstract

Muscle wasting accompanies diseases that are associated with chronic elevated levels of circulating inflammatory cytokines and oxidative stress. We previously demonstrated that tumor necrosis factor-α (TNF-α) inhibits myogenic differentiation via the activation of nuclear factor-κB (NF-κB). The goal of the present study was to determine whether this process depends on the induction of oxidative stress. We demonstrate here that TNF-α causes a decrease in reduced glutathione (GSH) during myogenic differentiation of C2C12 cells, which coincides with an elevated generation of reactive oxygen species. Supplementation of cellular GSH with N-acetyl-l-cysteine (NAC) did not reverse the inhibitory effects of TNF-α on troponin I promoter activation and only partially restored creatine kinase activity in TNF-α-treated cells. In contrast, the administration of NAC before treatment with TNF-α almost completely restored the formation of multinucleated myotubes. NAC decreased TNF-α-induced activation of NF-κB only marginally, indicating that the redox-sensitive component of the inhibition of myogenic differentiation by TNF-α occurred independently, or downstream of NF-κB. Our observations suggest that the inhibitory effects of TNF-α on myogenesis can be uncoupled in a redox-sensitive component affecting myotube formation and a redox independent component affecting myogenic protein expression.