The phosphoenolpyruvate carboxykinase (PEPCK) inhibitor, 3-mercaptopicolinic acid (3-MPA), induces myogenic differentiation in C2C12 cells.
The phosphoenolpyruvate carboxykinase (PEPCK) inhibitor, 3-mercaptopicolinic acid (3-MPA), induces myogenic differentiation in C2C12 cells.
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磷酸烯醇丙酮酸羧基酶（PEPCK）抑制剂3-羟基丙氨酸酸（3-MPA）诱导C2C12细胞中的肌生成分化。
DOI：
10.1038/s41598-020-79324-9
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发表时间：
2020-12-17
影响因子：
4.6
通讯作者：
Brameld JM
中科院分区：
文献类型：
--
作者：
Brearley MC;Daniel ZCTR;Loughna PT;Parr T;Brameld JM
磷酸烯醇式丙酮酸羧激酶（PEPCK）是一种具有胞浆（Pck 1/PEPCK-C）和线粒体（Pck 2/PEPCK-M）同种型的促凋亡酶。在这里，我们研究了PEPCK抑制剂3-巯基吡啶甲酸（3-MPA）对C2 C12肌细胞的影响。我们报告说，在C2 C12肌形成过程中，Pck 2 mRNA比Pck 1高50-5000倍，表明Pck 2是主要的PEPCK亚型。C2 C12细胞的增殖受到抑制，在剂量依赖性的方式后48小时3-MPA处理（0.01-1 mM）。3-MPA处理后C2 C12肌源性分化明显诱导（0.25、0.5、1 mM），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亚型（由Myh 4编码）是肥大的指示物，并且猪MYH 4启动子活性和内源性Myh 4 mRNA也被显著诱导。从第0天到第4天处理后，高剂量（0.5和/或1 mM）的3-MPA降低了Pck 2和与丝氨酸生物合成相关的基因（磷酸甘油酸脱氢酶，Phgdh;磷酸丝氨酸氨基转移酶-1，Psat 1）的mRNA表达。综上所述，由于Pck 2/PEPCK-M是C2 C12细胞中的主要亚型，我们推测3-MPA通过抑制PEPCK-M促进肌源性分化。然而，我们无法证实3-MPA抑制PEPCK-M酶活性，因为3-MPA干扰PEPCK酶测定，特别是在0.5和1 mM时。
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.
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影响因子：
3.7
作者：
Brown DM;Jones S;Daniel ZCTR;Brearley MC;Lewis JE;Ebling FJP;Parr T;Brameld JM
通讯作者：
Brameld JM
影响因子：
4.6
作者：
Brown DM;Williams H;Ryan KJ;Wilson TL;Daniel ZC;Mareko MH;Emes RD;Harris DW;Jones S;Wattis JA;Dryden IL;Hodgman TC;Brameld JM;Parr T
通讯作者：
Parr T
影响因子：
2.9
作者：
Balan, Marc D.;Mcleod, Matthew J.;Holyoak, Todd
通讯作者：
Holyoak, Todd
影响因子：
16
作者：
Vincent, Emma E.;Sergushichev, Alexey;Jones, Russell G.
通讯作者：
Jones, Russell G.
影响因子：
4.3
作者：
Szoka, Lukasz;Karna, Ewa;Palka, Jerzy
通讯作者：
Palka, Jerzy