Activation of hypoxia-inducible factor 1 in skeletal muscle cells after exposure to damaged muscle cell debris

Shock. 2011 Jun;35(6):632-8. doi: 10.1097/SHK.0b013e3182111f3d.


Skeletal muscle damage provokes complex repair mechanisms including recruitment of leukocytes as well as activation of myogenic precursor cells such as satellite cells. To study muscle cell repair mechanisms after muscle fiber damage, we used an in vitro model of scrape-injured myotubes. Exposing vital C2C12 myoblasts and myotubes to cell debris of damaged myotubes revealed mRNA upregulation of adrenomedullin (ADM), insulin-like growth factors 1 and 2, metallopeptidase 9, and monocyte chemoattractant protein11. When cell debris was treated with ultrasound, frozen in liquid nitrogen, or heat inactivated before addition to C2C12 cells, gene expression was drastically reduced or completely absent. Moreover, incubations of myoblasts with debris separated by transwell inserts indicated that direct cell contact is required for gene induction. Incubation with albumin and PolyIC ruled out that ADM induction by cell debris simply results from increased protein or nucleic acid concentrations in the supernatant. Because the genes, which were upregulated by cell debris, are potential target genes of hypoxia-inducible factor (HIF), cells were analyzed for HIF-1α expression. Western blot analysis showed accumulation of the α-subunit upon contact to cell debris. Knockdown of HIF-1α in C2C12 cells proved that activation of HIF-1 in response to cell debris was responsible for upregulating ADM and monocyte chemoattractant protein 1. Furthermore, by incubating cells on gas-permeable culture dishes, we excluded a reduced pericellular pO2 induced by cell debris as the cause for ADM upregulation. Our data suggest that damaged myofibers activate HIF-1 in neighboring myotubes and precursor myoblasts by direct contact, concomitantly upregulating factors necessary for angiogenesis, tissue regeneration, and phagocyte recruitment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenomedullin / genetics
  • Adrenomedullin / metabolism
  • Animals
  • Cell Line
  • Gene Expression / drug effects
  • Hypoxia-Inducible Factor 1 / biosynthesis*
  • Hypoxia-Inducible Factor 1, alpha Subunit / biosynthesis
  • Insulin-Like Growth Factor I / metabolism
  • Mice
  • Muscle Fibers, Skeletal / metabolism
  • Muscle, Skeletal / injuries*
  • Muscle, Skeletal / metabolism*
  • Muscle, Skeletal / physiology
  • Myoblasts / metabolism
  • RNA, Messenger / metabolism
  • Reactive Oxygen Species / metabolism
  • Regeneration


  • Hypoxia-Inducible Factor 1
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • RNA, Messenger
  • Reactive Oxygen Species
  • Adrenomedullin
  • Insulin-Like Growth Factor I