Myostatin inhibits myogenesis and promotes adipogenesis in C3H 10T(1/2) mesenchymal multipotent cells
Myostatin inhibits myogenesis and promotes adipogenesis in C3H 10T(1/2) mesenchymal multipotent cells
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DOI：
10.1210/en.2005-0362
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发表时间：
2005-08-01
期刊：
影响因子：
4.8
通讯作者：
Gonzalez-Cadavid, NF
中科院分区：
文献类型：
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作者：
Artaza, JN;Bhasin, S;Gonzalez-Cadavid, NF
哺乳动物肌肉生长抑制素基因的失活突变与肌肉质量增加和脂肪质量减少相关;相反，在骨骼肌中过表达肌肉生长抑制素的肌肉生长抑制素转基因小鼠具有减少的肌肉质量和增加的脂肪质量。我们研究了重组肌生成抑制素蛋白和抗肌生成抑制素抗体对间充质多能细胞成肌和成脂分化的影响。因此，将10 T（1/2）细胞与5 ′-氮杂胞苷孵育3天以诱导分化，然后用肌生长抑制素（Mst）羧基末端113个氨基酸的重组蛋白或多克隆抗Mst抗体处理3、7和14天。将表达全长375-氨基酸蛋白质的Mst cDNA质粒（pcDNA-Mst 375）和Mst沉默RNA（pSil-Mst）或随机序列（pSil-RS）共转染细胞3或7 d，并测定Mst表达。通过油红O染色前后脂肪细胞的定量图像分析、脂联素的免疫细胞化学和CCAAT/增强子结合蛋白-α的Western印迹来评估脂肪生成。通过定量图像分析-MyoD（Myo分化蛋白）、肌细胞生成素和肌球蛋白重链II型的免疫细胞化学，或通过肌细胞生成素的蛋白质印迹，估计肌细胞生成。5 '-氮杂胞苷介导的分化诱导内源性全长Mst表达。重组Mst羧基末端113个氨基酸抑制早期和晚期标志物的肌生成和刺激早期和晚期标志物的脂肪生成，而抗体对Mst发挥相反的效果。转染pcDNA-Mst 375后第7天，Myogenin的表达水平降低，而pSil-Mst的表达水平升高，有效地阻断了Mst 375的表达。总之，肌生长抑制素促进多能间充质细胞向成脂谱系分化并抑制肌生成。
Inactivating mutations of the mammalian myostatin gene are associated with increased muscle mass and decreased fat mass; conversely, myostatin transgenic mice that overexpress myostatin in the skeletal muscle have decreased muscle mass and increased fat mass. We investigated the effects of recombinant myostatin protein and antimyostatin antibody on myogenic and adipogenic differentiation of mesenchymal multipotent cells. Accordingly, 10T(1/2) cells were incubated with 5'-azacytidine for 3 d to induce differentiation and then treated with a recombinant protein for myostatin (Mst) carboxy terminal 113 amino acids or a polyclonal anti-Mst antibody for 3, 7, and 14 d. Cells were also cotransfected with a Mst cDNA plasmid expressing the full-length 375-amino acid protein (pcDNA-Mst375) and the silencer RNAs for either Mst (pSil-Mst) or a random sequence (pSil-RS) for 3 or 7 d, and Mst expression was determined. Adipogenesis was evaluated by quantitative image analysis of fat cells before and after oil-red-O staining, immunocytochemistry of adiponectin, and Western blot for CCAAT/enhancer binding protein-alpha. Myogenesis was estimated by quantitative image analysis-immunocytochemistry for MyoD (Myo differentiation protein), myogenin, and myosin heavy chain type II, or by Western blot for myogenin. 5'-azacytidine-mediated differentiation induced endogenous full-length Mst expression. Recombinant Mst carboxy terminal 113 amino acids inhibited both early and late markers of myogenesis and stimulated both early and late markers of adipogenesis, whereas the antibody against Mst exerted the reverse effects. Myogenin levels at 7 d after transfection of pcDNA-Mst375 were reduced as expected and elevated by pSil-Mst, which blocked efficiently Mst375 expression. In conclusion, myostatin promotes the differentiation of multipotent mesenchymal cells into the adipogenic lineage and inhibits myogenesis.