谷氨酰胺补充和电刺激对C2C12肌管蛋白合成通路的影响

发布时间：2018-05-14 06:25

本文选题：谷氨酰胺补充 + mTOR　；参考：《中国体育科技》2017年01期

【摘要】：目的:C2C12肌管为细胞模型,探讨电刺激模拟运动与谷氨酰胺补充对C2C12肌管蛋白质含量及mTOR通路的影响,进一步探明谷氨酰胺补充与蛋白质代谢之间的关系。方法:谷氨酰胺孵育分化5天的肌管48h后,给予一次电刺激(15 V,30ms,3Hz,90min)。分别在电刺激后0h、6h、18 h和24 h收样。实验共10组,单独对照组(Con);单独谷氨酰胺组(Gln);单独电刺激后不同时间点取样组(Es0、Es6、Es18、Es24);谷氨酰胺孵育加电刺激后不同时间点取样组(Gln+Es0、Gln+Es6、Gln+Es18、Gln+Es24)。BCA法测定各组肌管蛋白含量;RT-PCR法测定各组mTOR mRNA表达量及其下游因子S6K1 mRNA表达量。结果:1)与Con相比,Gln组的蛋白含量、mTOR及S6K1 mRNA均显著升高(P0.05)。2)单独电刺激(Es)后,随收样时间延长,肌管蛋白含量呈现先下降后上升趋势。Es6极显著下降(P0.01),Es18极显著上升(P0.01)。而mTOR和S6K1 mRNA的表达没有下降,呈现先上升后恢复正常的趋势。3)谷氨酰胺和电刺激(Gln+Es)联合作用后,无论是蛋白含量,还是mTOR及S6K1 mRNA表达,具有与单独电刺激相似的作用趋势,且Gln+Es各组数值均高于Es各组(P0.05)。此外,Es组mTOR表达在电刺激后18h达高峰,而Gln+Es组在6h达高峰。结论:1)单独电刺激、单独补充谷氨酰胺、两者的联合作用,均可通过mTOR通路中mTOR mRNA和S6K1 mRNA表达上升,增加蛋白合成,促进蛋白含量上升及肌管恢复。2)谷氨酰胺使电刺激后C2C12肌管mTOR mRNA表达提前,S6K1mRNA表达稳定处于高水平。
[Abstract]:Objective to investigate the effects of simulated exercise and glutamine supplementation on the protein content and mTOR pathway in C2C12 myotubes and to explore the relationship between glutamine supplementation and protein metabolism. Methods: after incubation with glutamine for 5 days, myotubes were incubated for 48 hours. The samples were collected at 0 h, 6 h, 18 h and 24 h after electrical stimulation, respectively. There were 10 groups, The content of myotube protein in each group was determined by RT-PCR at different time points after single electrical stimulation. The content of myotube protein was determined by RT-PCR in the sampling group at different time points after incubation with glutamine and at different time points after electric stimulation. The content of myotube protein in each group was determined by RT-PCR with the method of Gln Es0Gln Es6Gln Es18Gln Es24).BCA after glutamine incubation and electrical stimulation. The expression of mTOR mRNA and its downstream factor S6K1 mRNA. Results compared with Con, the protein content of Con group was significantly higher than that of Con group (P 0.05N 路2). With the prolongation of collecting time, the content of myotube protein decreased first and then increased. Es6 decreased significantly (P 0.01) and Es18 increased significantly (P 0.01). However, the expression of mTOR and S6K1 mRNA did not decrease, showing a tendency of rising first and then returning to normal. After the combination of glutamine and Glutamine, the protein content, mTOR and S6K1 mRNA expression were similar to those of electrical stimulation alone. The values of Gln es in each group were higher than that in es group (P 0.05). In addition, the expression of mTOR reached the peak at 18 h after electrical stimulation and reached the peak at 6 h in Gln es group. ConclusionThe combined effects of single electrical stimulation and glutamine supplementation can increase the expression of mTOR mRNA and S6K1 mRNA in the mTOR pathway and increase the protein synthesis. Glutamine promoted the increase of protein content and the recovery of myotube. 2) Glutamine stabilized the expression of S6K1 mRNA at a high level after electrical stimulation in the myotube of C2C12.
【作者单位】：暨南大学体育学院;华南师范大学体育科学学院;
【分类号】：G804.32

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