摘要
探究长时间不同频率EPS对C2C12肌管细胞代谢指标和基因表达的影响,试图建立EPS诱发C2C12肌管细胞TG堆积和线粒体功能增强的细胞模型。方法:采用4种EPS方案诱发C2C12肌管细胞收缩,实验分为5组:对照组(C组)、30 V+2 ms+0.5 Hz组(0.5 Hz组)、30 V+2 ms+1 Hz组(1 Hz组)、30 V+2 ms+5 Hz组(5 Hz组)、30 V+2 ms+10 Hz组(10 Hz组)。比色法检测细胞内TG、乳酸和ATP含量,RT-PCR方法检测细胞中IL-6、MyHCⅠ、MyHCⅡa、MyHCⅡb、MyHCⅡx、Cyt C和PGC-1α等mRNA含量。结果:1)与C组相比,EPS后即刻,频率0.5~10 Hz组乳酸含量非常显著增加(P<0.01);频率1 Hz和10 Hz组细胞中ATP含量均显著降低(P<0.05),其中频率0.5 Hz和5 Hz组细胞中ATP含量非常显著降低(P<0.01)。2)与C组相比,频率0.5 Hz和1 Hz组MyHCⅠ和MyHCⅡb的mRNA含量显著增加(P<0.05),频率5 Hz组MHC4种亚型mRNA含量变化不明显(P>0.05),频率10 Hz组MyHCⅠ、MyHCⅡa和MyHCⅡb亚型mRNA含量非常显著降低(P<0.01),而MyHCⅡx亚型mRNA含量非常显著增加(P<0.01)。3)与C组相比,频率0.5~10 Hz组PGC-1α和Cyt C mRNA含量均显著增加(P<0.05),同时EPS增加了C2C12细胞中IL-6 mRNA的含量(P<0.05)。4)与C组相比,4 d EPS方案停止后12 h,各组肌管细胞中TG含量均没有出现变化;4 d EPS方案停止后24 h,0.5 Hz组和1 Hz组肌管细胞中TG含量增加但无显著性(P>0.05);4 d EPS方案停止后36 h,0.5 Hz组和5 Hz组细胞中TG含量显著增加(P<0.05)。结论:EPS方案(电压14 V、频率1 Hz、单次时长2 ms和每天2 h共2 d,紧接着,电压30 V、频率0.5 Hz或1 Hz或5 Hz、单次时长2 ms和每天3 h共2 d,EPS停止后36 h)能够明显诱发C2C12肌管细胞TG含量堆积和线粒体功能增强。
Objective:To explore the effect of long time and different frequency electrical stimulation on the metabolic index and gene expression in C2C12 myotubes,so as to establish a cell model for EPS-induced TG accumulation and the increase of mitochondrial function in C2C12 myotubes.Methods:Four protocols of EPS were used to induce contraction of C2C12 myotubes.The experiment was divided into five groups:control group(C group),30V 2ms 0.5Hz group(0.5Hz group),30V 2ms 1Hz group(1Hz group),30V 2ms 5Hz.Group(5Hz group),30V 2ms 10Hz group(10Hz group).Colorimetric assay was used to detect intracellular TG,lactate,and ATP content.RT-PCR was used to detect mRNA levels of IL-6,MyHCⅠ,MyHCⅡa,MyHCⅡb,MyHCⅡx,Cyt C,and PGC-1α.Results:1)Compared with C group,the lactate content in the 0.5Hz-10Hz group was significantly increased(P<0.01)immediately after stimulation,and the ATP content in cells was significantly decreased in the 1Hz and 10Hz groups(P<0.05).However,the ATP content in cells at 0.5 Hz and 5 Hz groups significantly decreased(P<0.01).2)Compared with C group,MyHCⅠand MyHCⅡb were significantly increased in the 0.5 Hz/1 Hz group(P<0.05),and the four myosin heavy chain isoforms exhibited no change in 5 Hz group(P>0.05).The mRNA content of MyHCⅠ,MyHCⅡa,and MyHCⅡb subtypes was significantly decreased(P<0.01)in the 10 Hz group,while the mRNA levels of MyHCⅡx isoform was significantly increased(P<0.01).3)Compared with C group,the mRNA content of PGC-1αand Cytochrome C were significantly increased(P<0.05)in 0.5-10 Hz group,and the mRNA content of IL-6 was significantly increased by EPS in C2C12 cells(P<0.05).4)Compared with C group,TG content in EPS groups did not change in 12h after 4d electrical stimulation,and the TG content in 0.5Hz and 1Hz groups were increased in 24h after 4d electrical stimulation but there was no significant difference(P>0.05);however,the TG content in 0.5 Hz and 5 Hz groups were significantly increased in 36h after 4d electrical stimulation(P<0.05).Conclusion:the EPS protocols(Voltage
作者
孙景权
上官若男
李松波
严翊
谢敏豪
SUN Jingquan;SHANG GUAN Ruonan;LI Songbo;YAN Yi;XIE Minhao(Beijing Sport University,Beijing 100084,China;School of Physical Education,Sichuan University,Chengdu,Sichuan 610065,China;College of Physical Education,Chengdu University,Chengdu,Sichuan 610106,China;Institute of Sports Medicine,General Administration of Sports,Beijing 100061,China)
出处
《首都体育学院学报》
CSSCI
北大核心
2019年第5期459-466,共8页
Journal of Capital University of Physical Education and Sports
基金
国家自然科学基金青年项目(31801002)
中央高校基本科研业务费(2019自研-体育01)
