College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, Shanxi, China.
Department of Nephrology, Shanghai General Hosptial, Shanghai Jiaotong University, No. 100, Haining Road, Shanghai 200080, China.
Acta Histochem. 2019 Jul;121(5):539-545. doi: 10.1016/j.acthis.2019.04.001. Epub 2019 Apr 30.
This study investigated the effects of a CoCl-simulated hypoxic environment on the muscle fiber switching signaling pathways calcineurin A/nuclear factor of activated T cells cytoplasmic 1 (CnA/NFATc1) and myostatin. In this study, C2C12 muscle cells were cultured in vitro under CoCl-simulated chemical hypoxic conditions, the expression levels of CnA and myostatin were detected through qRT-PCR and Western blot analyses, and a positioning study of NFATc1 was carried out by immunofluorescence labeling. Results showed that CoCl treatment significantly increased the expression levels of CnA and myostatin. Moreover, the position of NFATc1 expression changed; actually, its expression in the nucleus considerably increased. Furthermore, CoCl-induced hypoxia inhibited the differentiation of C2C12 cells and reduced the expression levels of many slow- and fast-twitch muscles marker genes, but immunofluorescence staining results showed that the proportion of MyHC I type muscle fiber increased after CoCl treatment. The hypoxic environment simulated by CoCl can activate the signaling pathways CnA/NFATc1 and myostatin and increases the proportion of MyHC I type muscle fibers.
本研究探讨了 CoCl 模拟缺氧环境对肌纤维转换信号通路钙调神经磷酸酶 A/活化 T 细胞核因子细胞质 1(CnA/NFATc1)和肌肉生长抑制素的影响。在这项研究中,将 C2C12 肌肉细胞在体外进行 CoCl 模拟化学缺氧培养,通过 qRT-PCR 和 Western blot 分析检测 CnA 和肌肉生长抑制素的表达水平,并通过免疫荧光标记进行 NFATc1 的定位研究。结果表明,CoCl 处理显著增加了 CnA 和肌肉生长抑制素的表达水平。此外,NFATc1 的表达位置发生改变;实际上,其在细胞核中的表达显著增加。此外,CoCl 诱导的缺氧抑制了 C2C12 细胞的分化,并降低了许多慢肌和快肌标记基因的表达水平,但免疫荧光染色结果显示,CoCl 处理后 MyHC I 型肌纤维的比例增加。CoCl 模拟的缺氧环境可以激活 CnA/NFATc1 和肌肉生长抑制素信号通路,并增加 MyHC I 型肌纤维的比例。