Contractile C2C12 myotube model for studying exercise-inducible responses in skeletal muscle
Contractile C2C12 myotube model for studying exercise-inducible responses in skeletal muscle
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DOI：
10.1152/ajpendo.90280.2008
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发表时间：
2008-11-01
影响因子：
5.1
通讯作者：
Kanzaki, Makoto
中科院分区：
文献类型：
--
作者：
Nedachi, Taku;Fujita, Hideaki;Kanzaki, Makoto
作者：Jiang Jiang，Jiang Jiang，神崎Jiang用于研究骨骼肌运动诱导反应的收缩C2 C12肌管模型。Am J Physiol Endocrinol Metab 295：E1191-E1204，2008.首次发表于2008年9月9日; doi：10.1152/ajpendo.90280.2008。充足的运动导致骨骼肌以及其他组织/器官的各种生理变化，并且还负责维持健康肌肉，其通过GLUT 4易位显示增强的胰岛素响应性葡萄糖摄取。我们通过用电脉冲刺激（EPS）操纵细胞内Ca 2+瞬变产生高度发达的收缩性C2 C12肌管，EPS被赋予与体内骨骼肌相似的性质：1）由于从头肌节形成而引起的兴奋诱导的收缩活性，2）AMP激酶和应激激活的MAP激酶级联的激活，和3）通过GLUT 4再循环评估的改善的胰岛素反应性。Tbc 1d 1是一种与运动诱导的骨骼肌GLUT 4易位有关的Rab-GAP，在EPS诱导的收缩后，Tbc 1d 1也似乎在Ser（231）上磷酸化。此外，开关肌球蛋白重链（MHC）的表达从“快型”到“慢型”被观察到在C2 C12肌管赋予EPS诱导的重复收缩。利用这些高度发达的收缩性C2 C12肌管，我们确定了肌管衍生因子对EPS诱发的收缩反应，包括CXC趋化因子CXCL 1/KC和CXCL 5/LIX，以及IL-6，先前报道在体内收缩肌肉中上调。重要的是，动物跑步机实验表明，运动显著增加了CXCL 1/KC的全身水平，可能来自收缩肌肉。总之，这些结果证实，我们已经建立了一个专门的肌肉细胞培养模型，允许收缩诱导的细胞反应进行探索。利用这个模型，我们确定了收缩诱导型肌因子可能与运动诱导的代谢改变，免疫反应和血管生成有关。
Nedachi T, Fujita H, Kanzaki M. Contractile C2C12 myotube model for studying exercise-inducible responses in skeletal muscle. Am J Physiol Endocrinol Metab 295: E1191-E1204, 2008. First published September 9, 2008; doi:10.1152/ajpendo.90280.2008.-Adequate exercise leads to a vast variety of physiological changes in skeletal muscle as well as other tissues/organs and is also responsible for maintaining healthy muscle displaying enhanced insulin-responsive glucose uptake via GLUT4 translocation. We generated highly developed contractile C2C12 myotubes by manipulating intracellular Ca2+ transients with electric pulse stimulation (EPS) that is endowed with properties similar to those of in vivo skeletal muscle in terms of 1) excitation-induced contractile activity as a result of de novo sarcomere formation, 2) activation of both the AMP kinase and stress-activated MAP kinase cascades, and 3) improved insulin responsiveness as assessed by GLUT4 recycling. Tbc1d1, a Rab-GAP implicated in exercise-induced GLUT4 translocation in skeletal muscle, also appeared to be phosphorylated on Ser(231) after EPS-induced contraction. In addition, a switch in myosin heavy-chain (MHC) expression from "fast type" to "slow type" was observed in the C2C12 myotubes endowed with EPS-induced repetitive contractility. Taking advantage of these highly developed contractile C2C12 myotubes, we identified myotube-derived factors responsive to EPS-evoked contraction, including the CXC chemokines CXCL1/KC and CXCL5/LIX, as well as IL-6, previously reported to be upregulated in contracting muscles in vivo. Importantly, animal treadmill experiments revealed that exercise significantly increased systemic levels of CXCL1/KC, perhaps derived from contracting muscle. Taken together, these results confirm that we have established a specialized muscle cell culture model allowing contraction-inducible cellular responses to be explored. Utilizing this model, we identified contraction-inducible myokines potentially linked to the metabolic alterations, immune responses, and angiogenesis induced by exercise.