Production of interleukin-6 by skeletal myotubes: role of reactive oxygen species

Am J Respir Cell Mol Biol. 2002 May;26(5):587-93. doi: 10.1165/ajrcmb.26.5.4598.


In the present study we have tested the ability of reactive oxygen species (ROS) to stimulate the production of interleukin (IL)- 6 from skeletal myocytes. Differentiated C2C12 murine skeletal muscle cells (myotubes) exposed to pyrogallol (PYR), xanthine/ xanthine-oxidase (X/XO), or H(2)O(2) for 24 h exhibited a concentration-dependent increase in IL-6 production. Unlike myotubes, incubation of myoblasts and endothelial cells with X/XO or PYR did not result in increased IL-6 release. In myotubes, superoxide dismutase and catalase blocked the ROS-induced IL-6 release. Exposure of myotubes to H(2)O(2) increased steady-state IL-6 mRNA levels, and pretreatment of myotubes with actinomycin D or cycloheximide abolished the ROS-induced IL-6 production. In addition, pretreatment of cells with N-acetyl-cysteine blocked tumor necrosis factor (TNF)-alpha-induced IL-6 release, suggesting that endogenously produced ROS participate in IL-6 production. Myotubes stimulated with H(2)O(2) exhibited increased I kappa B-alpha phosphorylation and degradation, and treatment of C2C12 with ROS-generating agents increased activator protein (AP)-1 and nuclear factor (NF)-kappa B-dependent promoter activity. Finally, preincubation of myotubes with the pharmacologic inhibitor of NF-kappa B, diethyldithiocarbamate, or transient transfection with an I kappa B-alpha mutant, inhibited the ROS-stimulated IL-6 release. In conclusion, ROS stimulate IL-6 production from skeletal myotubes in a manner that involves transcriptional activation of the IL-6 gene through an NF-kappa B-dependent pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Ditiocarb / pharmacology
  • Dose-Response Relationship, Drug
  • Enzyme Inhibitors / pharmacology
  • Hydrogen Peroxide / pharmacology
  • Interleukin-6 / genetics
  • Interleukin-6 / metabolism*
  • Mice
  • Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • Mitogen-Activated Protein Kinases / metabolism
  • Muscle, Skeletal / cytology
  • Muscle, Skeletal / drug effects
  • Muscle, Skeletal / metabolism*
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / metabolism
  • Oxidants / pharmacology
  • Oxidation-Reduction / drug effects
  • Pyrogallol / pharmacology
  • RNA, Messenger / metabolism
  • Reactive Oxygen Species / metabolism*
  • Reactive Oxygen Species / pharmacology
  • Xanthine / pharmacology
  • Xanthine Oxidase / pharmacology
  • p38 Mitogen-Activated Protein Kinases


  • Enzyme Inhibitors
  • Interleukin-6
  • NF-kappa B
  • Oxidants
  • RNA, Messenger
  • Reactive Oxygen Species
  • Pyrogallol
  • Xanthine
  • Ditiocarb
  • Hydrogen Peroxide
  • Xanthine Oxidase
  • Mitogen-Activated Protein Kinases
  • p38 Mitogen-Activated Protein Kinases