Effects Of N-Oleoyl Glycine On The Protein Synthesis Of C2C12 Myoblast

The lean percentage is an important economic trait for pig production, which mainly depends on the development of skeletal muscle. Therefore, the study about proliferation and differentiation of pig skeletal muscle has become a highlight research. Fatty acyl amino acids are endogenous compounds, synthesized from the dehydration of fatty acids and amino acids, in the brain, spinal cord, skin, lung, muscle and fat tissues. A large number of studies showed that fatty acyl amino acids play an important role of anti-inflammatory, cell proliferation and cell apoptosis, as well as 3T3-L1 cell adipogenesis.To identify the effects of fatty acyl amino acids on proliferation, differentiation and protein deposition, C2C12 cells were treated with different types and different doses of fatty acyl amino acids. The most potential fatty acyl amino acids, N-Oleoyl glycine(OLGly), was adopted for further study. In the experiment, cell proliferation(by cell counting kit-8) and differentiation(by creatine kinase activity) and protein synthesis(by total protein content and puromycin incorporation level) were detected. Moreover, the m RNA expression level of PCNA, p27, p21, Myf-5, My OG, My HC I, My HC IIa, My HC IIb and My HC IIx were tested by q PCR. The protein expression level of m TOR, ERK, S6, Fox O1, MAFBx, Mu RF1, My HC I, My HC IIa, My HC IIb and My HC IIx were tested by western blot.The results showed that, 1) Fatty acyl amino acids had no effect on the proliferation and differentiation of C2C12 cells. However, fatty acyl amino acids could significantly promote protein synthesis of C2C12 myotubes at certain doses(P<0.05). Among them, OLGly demonstrated the most effective on protein synthesis. 2 or 20 ?mol/L of OLGly could significantly enhanced the protein synthesis of C2C12 myotubes in dose and time-dependent manners(P<0.05). However, the same dose of the mixture of oleic acid and glycine had no significant effect on protein synthesis, which means the effect of OLGly on protein synthesis in C2C12 myotubes is not mediated by its degradation metabolites. 2) The protein expression of the p-ERK and p-S6 were significantly elevated in response to 20 ?mol/L OLGly treatment for 30 min and 60 min(P<0.05). However, both OLGly and the mixture of its motabolites, oleic acid and glycine, unable to alter the protein expression of p-m TOR. For the protein degradation, OLGly had no effects on the protein expression level of p-Fox O1, MAFBx and Mu RF1 and the m RNA expression level of MAFBx and Mu RF1. 3) 20 ?M OLGly could significant reduce the protein and m RNA expression level of My HC IIb and My HC IIx(P<0.05), while unalter that of My HC I and My HC IIa(P>0.05).In summary, our data confirmed that OLGly could promote protein synthesis, had no effect on the proliferation and differentiation of C2C12 cells. The upregulation of ERK and S6 might be involved in the process for OLGly induced protein synthesis. The present study provide the experimental basis for the development of novel regulators to promote skeletal muscle growth and protein deposition.