Role of l-carnitine and oleate in myogenic differentiation: implications for myofiber regeneration.

Purpose: Myogenic progenitors play a critical role in injury-induced myofiber regeneration. The purpose of this study was to characterize the effects of oleate and L-carnitine on the overall behavior of proliferating myogenic progenitors (myoblasts) and its link to the mitochondrial biogenic process.

Methods: C2C12 myoblasts were cultured either with no treatment, oleate, L-carnitine, or their mixture. Proliferating myoblasts were investigated under a phase-contrast microscope. Myonuclei and myosin heavy chain were stained with DAPI and MF20 antibody, respectively, in differentiated myotubes and visualized under florescence microscopy. Mitochondrial biogenic markers and porin were assessed by qRT-PCR or immunoblotting.

Results: Increased proliferation rate was observed in myoblasts conditioned with oleate or a mixture of oleate and L-carnitine in contrast to that in non-treated (NT) and L-carnitine-treated myoblasts. Myoblast viability was not statistically different among all tested groups. Fusion index and myotube width were greater in oleate- or L-carnitine-conditioned myotubes than those in NT myotubes, with the greatest effect seen in myotubes conditioned with the mixture. The gene expressions of Pgc1-α, Nrf1, and Tfam were the greatest in myotubes conditioned with the mixture, whereas the level of Ncor1 expression was lower compared to those of the other groups. Protein level of porin was the greatest in myotubes conditioned with the mixture, followed by that of individually treated myotubes with oleate and L-carnitine.

Conclusion: These results provide a critical piece of cellular evidence that combined treatment of oleate and L-carnitine could serve as a potential therapeutic strategy to facilitate biological activation of myogenic progenitors.