Electrical pulse stimulation of cultured human skeletal muscle cells as an in vitro model of exercise.
Electrical pulse stimulation of cultured human skeletal muscle cells as an in vitro model of exercise.
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DOI：
10.1371/journal.pone.0033203
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Aas V
中科院分区：
文献类型：
--
作者：
Nikolić N;Bakke SS;Kase ET;Rudberg I;Flo Halle I;Rustan AC;Thoresen GH;Aas V
体育锻炼导致骨骼肌的大量适应性反应，并在健康的生活方式中发挥核心作用。由于运动诱导主要的全身反应，潜在的细胞机制是很难在体内研究。因此，希望开发一种类似于在培养的人肌管中训练的体外模型。电脉冲刺激（EPS）施加到粘附的人肌管。测定细胞内ATP、磷酸肌酸（PCr）和乳酸含量。葡萄糖和油酸的代谢进行了研究，使用放射性标记的底物，基因表达进行了分析，使用实时RT-PCR。线粒体含量和功能分别通过活体成像和柠檬酸合酶活性测定来测量。通过电泳和免疫印迹评估蛋白质表达。高频率，急性EPS增加脱氧葡萄糖的摄取和乳酸的生产，而ATP和PCr的细胞含量下降。慢性，低频EPS通过增加葡萄糖代谢（摄取和氧化）和完全脂肪酸氧化来增加培养肌管的氧化能力。EPS处理后，细胞内丙酮酸脱氢酶复合物4（PDK 4）mRNA表达水平显著升高，同时细胞色素C、白细胞介素6（IL-6）和肉毒素棕榈酰转移酶B（CPT 1 B）mRNA表达水平也有升高趋势。MitoTracker®Red FM的强度在慢性低频EPS 48小时后加倍。在EPS处理的细胞中，慢纤维类型标记物（MHCI）的蛋白质表达增加。我们的研究结果表明，在体外EPS（急性，高频率以及慢性，低频率）的人肌管可用于研究运动的影响。
Physical exercise leads to substantial adaptive responses in skeletal muscles and plays a central role in a healthy life style. Since exercise induces major systemic responses, underlying cellular mechanisms are difficult to study in vivo. It was therefore desirable to develop an in vitro model that would resemble training in cultured human myotubes. Electrical pulse stimulation (EPS) was applied to adherent human myotubes. Cellular contents of ATP, phosphocreatine (PCr) and lactate were determined. Glucose and oleic acid metabolism were studied using radio-labeled substrates, and gene expression was analyzed using real-time RT-PCR. Mitochondrial content and function were measured by live imaging and determination of citrate synthase activity, respectively. Protein expression was assessed by electrophoresis and immunoblotting. High-frequency, acute EPS increased deoxyglucose uptake and lactate production, while cell contents of both ATP and PCr decreased. Chronic, low-frequency EPS increased oxidative capacity of cultured myotubes by increasing glucose metabolism (uptake and oxidation) and complete fatty acid oxidation. mRNA expression level of pyruvate dehydrogenase complex 4 (PDK4) was significantly increased in EPS-treated cells, while mRNA expressions of interleukin 6 (IL-6), cytochrome C and carnitin palmitoyl transferase b (CPT1b) also tended to increase. Intensity of MitoTracker®Red FM was doubled after 48 h of chronic, low-frequency EPS. Protein expression of a slow fiber type marker (MHCI) was increased in EPS-treated cells. Our results imply that in vitro EPS (acute, high-frequent as well as chronic, low-frequent) of human myotubes may be used to study effects of exercise.
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DOI：
10.1016/s0006-291x(03)01124-0
发表时间：
2003-07-18
影响因子：
3.1
作者：
Al-Khalili, L;Cartee, GD;Krook, A
通讯作者：
Krook, A
影响因子：
3.7
作者：
Burch N;Arnold AS;Item F;Summermatter S;Brochmann Santana Santos G;Christe M;Boutellier U;Toigo M;Handschin C
通讯作者：
Handschin C
影响因子：
4.5
作者：
Gaster, M;Handberg, A;Schroder, HD
通讯作者：
Schroder, HD
影响因子：
8.2
作者：
Eriksen, L.;Dahl-Petersen, I.;Dela, F.
通讯作者：
Dela, F.
影响因子：
3.3
作者：
Chow, Lisa S.;Greenlund, Laura J.;Nair, K. Sreekumaran
通讯作者：
Nair, K. Sreekumaran