electrical pulse stimulation of cultured human skeletal muscle cells as an in vitro model of exercise电脉冲刺激培养人类骨骼肌细胞作为体外模型的运动.pdfVIP

Electrical Pulse Stimulation of Cultured Human Skeletal Muscle Cells as an In Vitro Model of Exercise ˇ ´ 1 1 1 1,2 1 Natasa Nikolic *, Siril Skaret Bakke , Eili Tranheim Kase , Ida Rudberg , Ingeborg Flo Halle , Arild C. 1 1 2 Rustan , G. Hege Thoresen , Vigdis Aas 1 Department of Pharmaceutical Biosciences, School of Pharmacy, University of Oslo, Oslo, Norway, 2 Faculty of Health Sciences, Oslo and Akershus University College of Applied Sciences, Oslo, Norway Abstract Background and Aims: 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. Methods: 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. Results: 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 gluco