Upregulation of IL-6 mRNA by IL-6 in skeletal muscle cells: role of IL-6 mRNA stabilization and Ca2+-dependent mechanisms

Am J Physiol Cell Physiol. 2007 Sep;293(3):C1139-47. doi: 10.1152/ajpcell.00142.2007. Epub 2007 Jul 5.

Abstract

Skeletal muscle cells have been established as significant producers of IL-6 during exercise. This IL-6 production is discussed as one possible mediator of the beneficial effects of physical activity on glucose and fatty acid metabolism. IL-6 itself could be the exercise-related factor that upregulates and maintains its own production. We investigated this hypothesis and the underlying molecular mechanism in cultured C(2)C(12) cells. IL-6 led to a rapid and prolonged increase in IL-6 mRNA, which was also found in human myotubes. Because IL-6 has been shown to activate AMP-activated kinase (AMPK), we studied whether, in turn, activated AMPK induces IL-6 expression. Pharmacological activation of AMPK with 5-aminoimidazole-4-carboxamide-1-beta-4-ribofuranoside upregulated IL-6 mRNA expression, which was blocked by knockdown of AMPK alpha(1) and alpha(2) using small, interfering RNA (siRNA) oligonucleotides. However, the effect of IL-6 was shown to be independent of AMPK, since the siRNA approach silencing the AMPK alpha-subunits did not reduce the upregulation of IL-6 induced by IL-6 stimulation. The self-stimulatory effect of IL-6 partly involves a Ca(2+)-dependent pathway: IL-6 increased intracellular Ca(2+), and intracellular blockade of Ca(2+) with a Ca(2+) chelator reduced the IL-6-mediated increase in IL-6 mRNA levels. Moreover, inhibition of Ca(2+)/calmodulin-dependent kinase kinase with STO-609 or the siRNA approach decreased IL-6 mRNA levels of control and IL-6-stimulated cells. A major, STO-609-independent mechanism is the IL-6-mediated stabilization of its mRNA. The data suggest that IL-6 could act as autocrine factor upregulating its mRNA levels, thereby supporting its function as an exercise-activated factor in skeletal muscle cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AMP-Activated Protein Kinases
  • Aminoimidazole Carboxamide / analogs & derivatives
  • Aminoimidazole Carboxamide / pharmacology
  • Animals
  • Autocrine Communication / drug effects
  • Autocrine Communication / physiology
  • Calcineurin / metabolism
  • Calcium / metabolism*
  • Calcium-Calmodulin-Dependent Protein Kinase Kinase
  • Cell Line
  • Enzyme Activation / drug effects
  • Enzyme Activation / physiology
  • Gene Expression Regulation / physiology
  • Hypoglycemic Agents / pharmacology
  • Interleukin-6 / genetics*
  • Interleukin-6 / metabolism*
  • Mice
  • Multienzyme Complexes / metabolism
  • Muscle Fibers, Skeletal / cytology
  • Muscle Fibers, Skeletal / metabolism*
  • Protein Serine-Threonine Kinases / metabolism
  • RNA Stability / physiology*
  • RNA, Messenger / metabolism
  • Ribonucleotides / pharmacology
  • Signal Transduction / drug effects
  • Signal Transduction / physiology
  • Up-Regulation / physiology
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • Hypoglycemic Agents
  • Interleukin-6
  • Multienzyme Complexes
  • RNA, Messenger
  • Ribonucleotides
  • Aminoimidazole Carboxamide
  • PRKAA2 protein, human
  • Protein Serine-Threonine Kinases
  • Calcium-Calmodulin-Dependent Protein Kinase Kinase
  • p38 Mitogen-Activated Protein Kinases
  • AMP-Activated Protein Kinases
  • PRKAA1 protein, human
  • Calcineurin
  • AICA ribonucleotide
  • Calcium