Myology Laboratory, Institute of Biomedical Problems RAS, 123007 Moscow, Russia.
Institute of Molecular Biology and Genetics, Almazov National Medical Research Centre, 197341 Saint Petersburg, Russia.
Int J Mol Sci. 2022 Aug 15;23(16):9150. doi: 10.3390/ijms23169150.
The structure and function of soleus muscle fibers undergo substantial remodeling under real or simulated microgravity conditions. However, unloading-induced changes in the functional activity of skeletal muscle primary myoblasts remain poorly studied. The purpose of our study was to investigate how short-term and long-term mechanical unloading would affect cultured myoblasts derived from rat soleus muscle. Mechanical unloading was simulated by rat hindlimb suspension model (HS). Myoblasts were purified from rat soleus at basal conditions and after 1, 3, 7, and 14 days of HS. Myoblasts were expanded in vitro, and the myogenic nature was confirmed by their ability to differentiate as well as by immunostaining/mRNA expression of myogenic markers. The proliferation activity at different time points after HS was analyzed, and transcriptome analysis was performed. We have shown that soleus-derived myoblasts differently respond to an early and later stage of HS. At the early stage of HS, the proliferative activity of myoblasts was slightly decreased, and processes related to myogenesis activation were downregulated. At the later stage of HS, we observed a decrease in myoblast proliferative potential and spontaneous upregulation of the pro-myogenic program.
在真实或模拟微重力条件下,比目鱼肌纤维的结构和功能会发生显著重塑。然而,对于骨骼肌肉原代成肌细胞在负荷减少时的功能活动的变化,研究甚少。本研究旨在探讨短期和长期机械性失重在何种程度上影响大鼠比目鱼肌来源的培养成肌细胞。通过大鼠后肢悬吊模型(HS)模拟机械性失载。在基础条件下以及 HS 后 1、3、7 和 14 天时,从大鼠比目鱼肌中纯化成肌细胞。成肌细胞在体外扩增,并通过其分化能力以及成肌标志物的免疫染色/ mRNA 表达来确认其成肌特性。分析了 HS 后不同时间点的增殖活性,并进行了转录组分析。我们已经表明,比目鱼肌来源的成肌细胞对 HS 的早期和晚期阶段有不同的反应。在 HS 的早期阶段,成肌细胞的增殖活性略有下降,与肌生成激活相关的过程被下调。在 HS 的晚期阶段,我们观察到成肌细胞增殖潜能下降,原肌生成程序自发上调。