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心肌肌动蛋白α1对牛成肌细胞和前脂肪细胞增殖及分化的影响

Effect of Actin Alpha Cardiac Muscle 1 on the Proliferation and Differentiation of Bovine Myoblasts and Preadipocytes.

作者信息

Li Anqi, Su Xiaotong, Tian Yuan, Song Guibing, Zan Linsen, Wang Hongbao

机构信息

College of Animal Science and Technology, Northwest A&F University, Yangling, Xianyang 712100, China.

National Beef Cattle Improvement Centre, Yangling, Xianyang 712100, China.

出版信息

Animals (Basel). 2021 Dec 6;11(12):3468. doi: 10.3390/ani11123468.

DOI:10.3390/ani11123468
PMID:34944244
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8698029/
Abstract

Actin Alpha Cardiac Muscle 1 (ACTC1) gene is a differentially expressed gene screened through the co-culture system of myoblasts-preadipocytes. In order to study the role of this gene in the process of proliferation and differentiation of bovine myoblasts and preadipocytes, the methods of the knockdown, overexpression, and ectopic expression of were used in this study. After knockdown in bovine myoblasts and inducing differentiation, the sizes and numbers of myotube formation were significantly reduced compared to the control group, and myogenic marker genes-MYOD1, MYOG, MYH3, MRF4, MYF5, CKM and MEF2A-were significantly decreased ( < 0.05, < 0.01) at both the mRNA and protein levels of myoblasts at different differentiation stages (D0, D2, D4, D6 and D8). Conversely, overexpression induced the inverse result. After ectopic expression of in bovine preadipocytes and induced differentiation, the number and size of lipid droplets were significantly higher than those of the control group, and the expression of adipogenic marker genes-FABP4, SCD1, PPARγ and FASN-were significantly increased ( < 0.05, < 0.01) at the mRNA and protein levels of preadipocytes at different differentiation stages. Flow cytometry results showed that both the knockdown and overexpression of inhibited the normal cell cycle of myoblasts; however, ectopic expression of in adipocytes induced no significant cell cycle changes. This study is the first to explore the role of in bovine myogenesis and lipogenesis and demonstrates that promotes the differentiation of bovine myoblasts and preadipocytes, affecting the proliferation of myoblasts.

摘要

心肌肌动蛋白α1（ACTC1）基因是通过成肌细胞-前脂肪细胞共培养系统筛选出的差异表达基因。为了研究该基因在牛成肌细胞和前脂肪细胞增殖与分化过程中的作用，本研究采用了基因敲低、过表达和异位表达的方法。在牛成肌细胞中敲低该基因并诱导分化后，与对照组相比，肌管形成的大小和数量显著减少，并且在不同分化阶段（D0、D2、D4、D6和D8）的成肌细胞的mRNA和蛋白质水平上，成肌标记基因——肌分化抗原1（MYOD1）、肌细胞生成素（MYOG）、肌球蛋白重链3（MYH3）、肌肉调节因子4（MRF4）、肌因子5（MYF5）、肌酸激酶M型（CKM）和肌细胞增强因子2A（MEF2A）——均显著降低（P<0.05，P<0.01）。相反，该基因过表达则诱导出相反的结果。在牛前脂肪细胞中异位表达该基因并诱导分化后，脂滴的数量和大小显著高于对照组，并且在不同分化阶段的前脂肪细胞的mRNA和蛋白质水平上，脂肪生成标记基因——脂肪酸结合蛋白4（FABP4）、硬脂酰辅酶A去饱和酶1（SCD1）、过氧化物酶体增殖物激活受体γ（PPARγ）和脂肪酸合酶（FASN）——均显著增加（P<0.05，P<0.01）。流式细胞术结果表明，该基因的敲低和过表达均抑制了成肌细胞的正常细胞周期；然而，在脂肪细胞中异位表达该基因未诱导出显著的细胞周期变化。本研究首次探索了该基因在牛肌肉生成和脂肪生成中的作用，并证明该基因促进牛成肌细胞和前脂肪细胞的分化，影响成肌细胞的增殖。

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