Creatine supplementation prevents the inhibition of myogenic differentiation in oxidatively injured C2C12 murine myoblasts.
Creatine (Cr), one of the most popular nutritional supplements among athletes, has been recently shown to prevent the cytotoxicity caused by different oxidative stressors in various mammalian cell lines, including C2C12 myoblasts, via a direct antioxidant activity. Here, the effect of Cr on the differentiating capacity of C2C12 cells exposed to H(2)O(2) has been investigated. Differentiation into myotubes was monitored using morphological, ultrastructural, and molecular techniques. Treatment with H(2)O(2) (1 h) not only caused a significant (30%) loss of cell viability, but also abrogated the myogenic ability of surviving C2C12. Cr-supplementation (24 h prior to H(2)O(2) treatment) was found to prevent these effects. Interestingly, H(2)O(2)-challenged cells preconditioned with the established antioxidants trolox or N-acetyl-cysteine, although cytoprotected, did not display the same differentiating ability characterizing oxidatively-injured, Cr-supplemented cells. Besides acting as an antioxidant, Cr increased the level of muscle regulatory factors and IGF1 (an effect partly refractory to oxidative stress), the cellular availability of phosphocreatine and seemed to exert some mitochondrially-targeted protective activity. It is concluded that Cr preserves the myogenic ability of oxidatively injured C2C12 via a pleiotropic mechanism involving not only its antioxidant capacity, but also the contribution to cell energy charge and effects at the transcriptional level which common bona fide antioxidants lack.
Sestili
Piero
P
Dipartimento di Scienze Biomolecolari, Sezione di Scienze Motorie e della Salute, Università degli Studi di Urbino Carlo Bo, Urbino (PU), Italy. piero.sestili@uniurb.it
Barbieri
Elena
E
Martinelli
Chiara
C
Battistelli
Michela
M
Guescini
Michele
M
Vallorani
Luciana
L
Casadei
Lucia
L
D'Emilio
Alessandra
A
Falcieri
Elisabetta
E
Piccoli
Giovanni
G
Agostini
Deborah
D
Annibalini
Giosuè
G
Paolillo
Marco
M
Gioacchini
Anna Maria
AM
Stocchi
Vilberto
V
D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002374:Catalase / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003401:Creatine / Q000032:analysis / Q000494:pharmacology*; D019587:Dietary Supplements; D005979:Glutathione Peroxidase / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000235:genetics; D051379:Mice; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D010084:Oxidation-Reduction; D040901:Proteomics; D012333:RNA, Messenger / Q000032:analysis
D000975:Antioxidants; D012333:RNA, Messenger; D007334:Insulin-Like Growth Factor I; D000255:Adenosine Triphosphate; D006861:Hydrogen Peroxide; D002374:Catalase; D005979:Glutathione Peroxidase; D003401:Creatine