Skeletal muscle adaptation is mediated by cooperative regulation of metabolism, signal transduction, and gene expression. However, the global regulatory mechanism remains unclear. To address this issue, we performed electrical pulse stimulation (EPS) in differentiated C2C12 myotubes at low and high frequency, carried out metabolome and transcriptome analyses, and investigated phosphorylation status of signaling molecules. EPS triggered extensive and specific changes in metabolites, signaling phosphorylation, and gene expression during and after EPS in a frequency-dependent manner. We constructed trans-omic network by integrating these data and found selective activation of the pentose phosphate pathway including metabolites, upstream signaling molecules, and gene expression of metabolic enzymes after high-frequency EPS. We experimentally validated that activation of these molecules after high-frequency EPS was dependent on reactive oxygen species (ROS). Thus, the trans-omic analysis revealed ROS-dependent activation in signal transduction, metabolome, and transcriptome after high-frequency EPS in C2C12 myotubes, shedding light on possible mechanisms of muscle adaptation.

骨骼肌适应性是通过代谢，信号转导和基因表达的协同调节来介导的。但是，全球监管机制仍不清楚。为了解决这个问题，我们在低频和高频下对分化的C2C12肌管进行了电脉冲刺激（EPS），进行了代谢组和转录组分析，并研究了信号分子的磷酸化状态。EPS在EPS期间和之后以频率依赖的方式触发了代谢产物的广泛而特定的变化，信号磷酸化和基因表达。通过整合这些数据，我们构建了跨组学网络，并发现了磷酸戊糖途径的选择性激活，包括代谢产物，上游信号分子以及高频EPS后代谢酶的基因表达。我们通过实验验证了高频EPS后这些分子的激活取决于活性氧（ROS）。因此，转组学分析揭示了在C2C12肌管中高频EPS后，信号转导，代谢组和转录组中的ROS依赖性激活，从而阐明了肌肉适应的可能机制。