Creatine supplementation prevents the inhibition of myogenic differentiation in oxidatively injured C2C12 murine myoblasts

Abstract

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.

The Pleiotropic Effect of Physical Exercise on Mitochondrial Dynamics in Aging Skeletal Muscle

“…The overexpression of PGC-1 in the skeletal muscle of aged mice improves oxidative capacity, suppresses mitochondrial degradation, and prevents muscle atrophy [94]. Building on our group's previous research in this area [95,96], we have recently studied the role of PGC-1 in C2C12 myoblasts subjected to oxidative stress during the early stages of differentiation, as well as the effect of H 2 O 2 (0.3 mM) on PGC-1 expression and its relationship with AMPK activation. We found that 1 h treatment with H 2 O 2 causes mitochondrial fragmentation and a mild mitochondrial impairment with a consequent marked increase in PGC-1 mRNA expression, as described by Kang et al [97] and Irrcher et al [98].…”

Effects of dietary guanidinoacetic acid and betaine supplementation on performance, blood biochemical parameters and antioxidant status of broilers subjected to cold stress

“…Theoretically, if GAA metabolism remains undisturbed and/or low-to-modest quantities of exogenous GAA have been provided, a cumulative effect of the intervention might be neutral or antioxidant protection through creatineand arginine-driven medium. Creatine, the end product of GAA utilization, is thought to have antioxidant capacity in some studies (Sestili et al 2006;Sestili et al 2009), but reduce antioxidant status in others (Percário et al 2012). According to Fathi and Tanha (2015), arginine supplementation increased plasma GPx activity and decreased MDA level in the broiler with cold-induced acites.…”

The expression analysis of mouse interleukin-6 splice variants argued against their biological relevance

“…The murine C2C12 cells were cultured as previously described (23). During C2C12 differentiation, cells were harvested by trypsinisation in proliferative phase (myoblasts 60-70% confluence), upon reaching confluence (T0) and in the following four days of differentiation (T1, T2, T3 and T4).…”