磷酸烯醇丙酮酸羧激酶（PEPCK）抑制剂 3-巯基吡啶甲酸（3-MPA）可诱导 C2C12 细胞的成肌分化。
The phosphoenolpyruvate carboxykinase (PEPCK) inhibitor, 3-mercaptopicolinic acid (3-MPA), induces myogenic differentiation in C2C12 cells.
机构信息
School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire, LE12 5RD, UK.
Department of Medicine-Cardiology, University of California, Los Angeles, CA, USA.
出版信息
Sci Rep. 2020 Dec 17;10(1):22177. doi: 10.1038/s41598-020-79324-9.
Phosphoenolpyruvate carboxykinase (PEPCK) is a gluconeogenic enzyme with a cytosolic (Pck1/PEPCK-C) and mitochondrial (Pck2/PEPCK-M) isoform. Here we investigate the effect of 3-mercaptopicolinic acid (3-MPA), a PEPCK inhibitor, on C2C12 muscle cells. We report that Pck2 mRNA is 50-5000-fold higher than Pck1 during C2C12 myogenesis, indicating Pck2 is the predominant PEPCK isoform. C2C12 cell proliferation was inhibited in a dose-dependent manner following 48 h 3-MPA treatment (0.01-1 mM). C2C12 myogenic differentiation was significantly induced following 3-MPA treatment (0.25, 0.5, 1 mM) from day 0 of differentiation, demonstrated by increased creatine kinase activity, fusion index and myotube diameter; likewise, the myosin heavy chain (MyHC)-IIB isoform (encoded by Myh4) is an indicator of hypertrophy, and both porcine MYH4-promoter activity and endogenous Myh4 mRNA were also significantly induced. High doses (0.5 and/or 1 mM) of 3-MPA reduced mRNA expression of Pck2 and genes associated with serine biosynthesis (Phosphoglycerate dehydrogenase, Phgdh; phosphoserine aminotransferase-1, Psat1) following treatment from days 0 and 4. To conclude, as Pck2/PEPCK-M is the predominant isoform in C2C12 cells, we postulate that 3-MPA promoted myogenic differentiation through the inhibition of PEPCK-M. However, we were unable to confirm that 3-MPA inhibited PEPCK-M enzyme activity as 3-MPA interfered with the PEPCK enzyme assay, particularly at 0.5 and 1 mM.
磷酸烯醇式丙酮酸羧激酶（PEPCK）是一种糖异生酶，具有胞质（Pck1/PEPCK-C）和线粒体（Pck2/PEPCK-M）同工型。在这里，我们研究了 PEPCK 抑制剂 3-巯基吡啶甲酸（3-MPA）对 C2C12 肌肉细胞的影响。我们报告说，在 C2C12 成肌过程中，Pck2 mRNA 是 Pck1 的 50-5000 倍，表明 Pck2 是主要的 PEPCK 同工型。48 小时 3-MPA 处理（0.01-1 mM）后，C2C12 细胞增殖呈剂量依赖性抑制。从分化的第 0 天开始，3-MPA 处理（0.25、0.5、1 mM）显著诱导 C2C12 肌生成分化，表现为肌酸激酶活性、融合指数和肌管直径增加；同样，肌球蛋白重链（MyHC）-IIB 同工型（由 Myh4 编码）是肥大的指标，猪 MYH4-启动子活性和内源性 Myh4 mRNA 也显著增加。高剂量（0.5 和/或 1 mM）的 3-MPA 在从第 0 天和第 4 天开始处理后，降低了 Pck2 和与丝氨酸生物合成相关的基因（磷酸甘油酸脱氢酶，Phgdh；磷酸丝氨酸转氨酶-1，Psat1）的 mRNA 表达。总之，由于 Pck2/PEPCK-M 是 C2C12 细胞中的主要同工型，我们推测 3-MPA 通过抑制 PEPCK-M 促进肌生成分化。然而，我们无法确认 3-MPA 抑制了 PEPCK-M 酶活性，因为 3-MPA 干扰了 PEPCK 酶测定，特别是在 0.5 和 1 mM 时。
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