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Special Issue:
动物科学合辑Animal Science
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| C-type natriuretic peptide stimulates chicken myoblast differentiation through NPRB/NPRC receptors and metabolism pathway |
| HUANG Hua-yun1*, LIANG Zhong1*, LIU Long-zhou1, 2, LI Chun-miao1, HUANG Zhen-yang1, WANG Qian-bao1, LI Shou-feng1, ZHAO Zhen-hua1 |
1 Jiangsu Institute of Poultry Science, Yangzhou 225125, P.R.China
2 College of Animal Science, Yangtze University, Jingzhou 8060550, P.R.China
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摘要
【研究目的】肉鸡骨骼肌的发育情况与肉产量及质量密切相关，而骨骼肌的发育主要依赖于成肌细胞的分化。利钠肽家族成员在哺乳动物骨骼肌的肌管形成及脂肪氧化过程中起重要作用，而C-型利钠肽激素（CNP）作为利钠肽家族的重要成员之一，它对骨骼肌的发育调控作用及机制尚未见相关报道，为更好的理解利钠肽家族在骨骼肌发育中的作用及机制，本研究通过外源10-7 mol L-1CNP诱导鸡的原代成肌细胞，探讨CNP对鸡成肌细胞分化的作用及机制。【研究方法】利用CCK8和EDU染色法检测成肌细胞的增殖，Q-PCR技术检测相关基因的表达，转录组测序及生物信息学分析筛选成肌细胞中响应CNP调控的差异基因及显著富集的信号通路。【研究结果】结果表明：与对照组相较，外源添加CNP后，成肌细胞的增殖能力显著增强（P < 0.05）；成肌细胞分化的标志基因MYOD和MYOG表达显著上升（P < 0.05）；CNP的特异性受体NPRB（Natriuretic peptide receptor B） 和清除性受体NPRC（Natriuretic peptide receptor C）的表达显著上调（P < 0.05）；转录组测序分析结果表明，142个差异基因（上调基因84个，下调基因58个）响应CNP对成肌细胞分化的调控作用（P < 0.05）；142个差异表达基因显著富集在8个信号通路（P < 0.05），而代谢通路里富集了16个差异表达基因，其中phospholipase C delta 4（PLCD4）、phospholipase C beta 2（PLCB2）、phosphoglycerate mutase 1（PGAM1）、 creatine kinase B（CKB）、 peroxiredoxin 6（PRDX6）和 CD38这6个基因与骨骼肌的发育密切相关。【研究结论】综上所述，CNP通过上调NPRB/NPRC及富集在代谢通路里的关键基因（PLCD4、 PLCβ2、 PGAM1、 CKB、 PRDX6和 CD38）促进成肌细胞的分化。【创 新 性】本研究利用功能基因验证法和高通量转录组测序法相结合的方法，首次系统阐述了CNP对成肌细胞分化的调控作用及机制，为更好地理解利钠肽家族在骨骼肌发育中的作用及机制奠定了基础。
Abstract Skeletal muscle development is closely related with the amount of meat production and its quality in chickens. Natriuretic peptides (NPs) play an important role in myotube formation and fat oxidation of skeletal muscle in animals. The effect of C-type natriuretic peptide (CNP), an important member of the NPs, and its underlying molecular mechanisms in skeletal muscle are incompletely understood. Treatment of myoblasts with CNP led to enhanced proliferation/differentiation and significantly upregulated (P<0.05) mRNA expression of the CNP receptors natriuretic peptide receptor B (NPRB) and the clearance receptor C (NPRC). In cells exposed to CNP, 142 differentially expressed genes (84 up-regulation and 58 down-regulation) (P<0.05) were identified by RNA-sequencing compared with those in control cells. Sixteen genes were significantly enriched (P<0.05) in the metabolic pathway, and six of them (phospholipase C β4, phospholipase C β2, phosphoglycerate mutase 1, creatine kinase B, peroxiredoxin 6 and CD38) were closely related to skeletal muscle development and differentially expressed. In conclusion, CNP stimulated differentiation of myoblasts by upregulating expression of the NPRB and NPRC receptors and enriching key genes in the metabolic pathway.
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Received: 02 September 2020
Accepted: 03 April 2021
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| Fund: The research was supported by the Natural Science Foundation of Jiangsu Province, China (BK20191217), the Independent Innovation Fund Project of Agricultural Science and Technology of Jiangsu (CX(20)2012), the China Agriculture Research System of MOF and MARA (CARS-41-Z05), the Key Laboratory of Poultry Genetics and Breeding in Jiangsu (JQLAB-ZZ-201705 and JQLAB-ZZ-202008), and the Special Funds for Transformation of Scientific and Technological Projects in Jiangsu Province (BA2019049). |
| About author: Huang Hua-yun, E-mail: huanghuayun520@163.com; Correspondence Zhao Zhen-hua, Tel/Fax: +86-514-85599073, E-mail: zzh0514@163.com
* These authors contributed equally to this study. |
Cite this article:
HUANG Hua-yun, LIANG Zhong, LIU Long-zhou, LI Chun-miao, HUANG Zhen-yang, WANG Qian-bao, LI Shou-feng, ZHAO Zhen-hua.
2022.
C-type natriuretic peptide stimulates chicken myoblast differentiation through NPRB/NPRC receptors and metabolism pathway. Journal of Integrative Agriculture, 21(2): 496-503.
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