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Glutamine Enhances Protein Content by the mTOR Pathway in Electrical Stimulated C2C12 Myotubes.

Heling Dong, Hongyuan Wu, Jun Zhao, Ruizhang Lin, Yonghong Zhang, Xu Li, Yinwei Huang, Xiaoyang Xu

Mammalian target of rapamycin (mTOR) plays an important role in regulating protein synthesis. This paper discusses the effects of glutamine and electrical stimulation on protein content and the mTOR pathway in C2C12, providing theories on how glutamine improves sports performance and recovery. After 5 days of differentiation, C2C12 myotubes were incubated in 4 mM or 10 mM doses of glutamine for 48 hours, and then stimulated to contract for 90 mins at 15v, 3Hz, and 30ms. After electrical stimulation, samples of Es and Gln+Es were harvested at 0, 6, 18, and 24 hours. The BCA Protein Assay Kit was used to measure total protein content (μg/ cm²). The expression of mTOR and S6K1 were measured by RT-PCR. The expression of p-S6 and myosin heavy chain (MHC) were measured by western blot. The results showed that total protein content, mTOR, p-S6, MHC expression of Gln were greater than that in Con (P<0.05). Compared with the protein content found in Con, that of Es18 and Es24 increased significantly (P<0.01). When the expression of mTOR and p-S6 were compared, it was apparent that Gln+Es and Es had similar trends, which was a decline in protein followed by a subsequent rise. The difference was that Es reached the peak in 18 hours, while Gln+Es did so in 6 h. The MHC expression in both Es and Gln+Es changed stable with a similar trend, decline followed by rise. Electrical stimulation and glutamine promote expression of MHC through the mTOR pathway, increase protein synthesis, and promote the rise of protein content and muscle recovery.