A reducing redox environment promotes C2C12 myogenesis: implications for regeneration in aged muscle.
Intracellular redox potential of skeletal muscle becomes progressively more oxidized with aging, negatively impacting regenerative ability. We examined the effects of oxidizing redox potential on terminal differentiation of cultured C2C12 myoblasts. Redox potentials were manipulated by changing the culture O(2) environment, by free radical scavenging, or addition of H(2)O(2.) Intracellular reactive oxygen species (ROS) production was higher in 20% environmental O(2) and in this condition, redox potential became progressively oxidized compared to cultures in 6% O(2.) Treatment with a ROS trapping agent (phenyl-N-tert-butylnitrone, PBN) caused reducing redox potentials and enhanced C2C12 differentiation, while addition of 25 micromolar H(2)O(2) to cells in 20% O(2) dramatically slowed differentiation. Under these most oxidative conditions, quantitative PCR showed a significant decrease in myogenic basic helix-loop-helix transcription factor expression compared to cultures treated with PBN or grown in 6% O(2). Thus, oxidative intracellular environments impair myoblast differentiation, while reducing environments favor myogenesis.
Hansen
Jason M
JM
Department of Anesthesiology, School of Medicine, Emory University, 1462 Clifton Road NE, Suite 420, Atlanta, GA 30322, USA.
Klass
Markus
M
Harris
Craig
C
Csete
Marie
M
D000375:Aging / Q000187:drug effects / Q000502:physiology*; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D003545:Cysteine / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D016166:Free Radical Scavengers / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism; D051379:Mice; D024510:Muscle Development / Q000187:drug effects / Q000502:physiology*; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000187:drug effects; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D032448:Myoblasts, Skeletal / Q000166:cytology* / Q000187:drug effects; D018995:Myosin Heavy Chains / Q000378:metabolism; D010084:Oxidation-Reduction / Q000187:drug effects; D010100:Oxygen / Q000494:pharmacology; D010545:Peroxides / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D012038:Regeneration / Q000187:drug effects / Q000502:physiology*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D051792:Basic Helix-Loop-Helix Proteins; D053148:Caspase 3; D003545:Cysteine; D016166:Free Radical Scavengers; D005978:Glutathione; D018995:Myosin Heavy Chains; D010100:Oxygen; D010545:Peroxides; D017382:Reactive Oxygen Species; D048051:p38 Mitogen-Activated Protein Kinases