Damage to developing mouse skeletal muscle myotubes in culture: protective effect of heat shock proteins.
Damage to skeletal muscle occurs following excessive exercise, upon reperfusion following ischaemia and in disease states, such as muscular dystrophy. Key mechanisms by which damage is thought to occur include a loss of intracellular calcium homeostasis, loss of energy supply to the cell, increased activity of oxidising free radical-mediated reactions and activation of apoptosis pathways. An increased cellular content of heat shock proteins (HSPs) has been shown to protect skeletal muscle against some forms of damage, although the mechanistic basis of this protection is not clearly understood. The aim of this study was to establish a cell culture-based model of damage to C2C12 skeletal muscle cells using the calcium ionophore, A23187 and the mitochondrial uncoupler, 2,4-dinitrophenol (DNP) as damaging agents. Treatment of cells with 1 mM DNP for 60 min resulted in the release of 63.5 % of intracellular creatine kinase (CK) activity over the 3 h experimental period. Treatment of cells with 10 microM A23187 for 30 min resulted in the release of 47.9 % of CK activity. Exposure of myotubes to a period of hyperthermia resulted in a significant increase in their content of HSP25, HSP60, HSC70 (heat shock cognate) and HSP70. This increase in HSPs was associated with significant protection against both DNP-induced and A23187-induced damage to the myotubes. These results indicate that an increased content of HSPs may provide protection against the muscle damage that occurs by a pathological increase in intracellular calcium or uncoupling of the mitochondrial respiratory chain.
Maglara
A A
AA
Department of Medicine, University of Liverpool, Liverpool L69 3GA, UK.
Vasilaki
A
A
Jackson
M J
MJ
McArdle
A
A
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018834:Chaperonin 60 / Q000502:physiology; D003080:Cold Temperature; D003402:Creatine Kinase / Q000378:metabolism; D004734:Energy Metabolism; D018840:HSP70 Heat-Shock Proteins / Q000502:physiology; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000502:physiology*; D006706:Homeostasis; D007700:Kinetics; D051379:Mice; D008954:Models, Biological; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000473:pathology* / Q000502:physiology; D018482:Muscle, Skeletal / Q000187:drug effects / Q000473:pathology* / Q000502:physiology
D018834:Chaperonin 60; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D000001:Calcimycin; D003402:Creatine Kinase; D002118:Calcium