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GH蛋白诱导鸡原代成肌细胞表达IGF1条件优化 被引量:1

The Effect of GH Protein on the Expression of IGF1 in Chicken (Gallus gallus) Myoblast Cells
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摘要 生长激素(growth hormone, GH)具有促进动物生长发育的作用,其通过内分泌系统与生长激素受体(growth hormone receptor, GHR)结合形成GH-GHR-IGF1信号通路,在组织和器官发育过程中发挥功能。胰岛素样生长因子1(insulin-like growth factor 1, IGF1)是一种生长调节多肽,对动物的生长发育和生理调节有重要作用。为了确定体外培养条件下GH诱导鸡成肌细胞表达IGF1的最佳添加方式和浓度,本实验通过分离培养鸡(Gallus gallus)原代成肌细胞,研究GH对下游IGF1 mRNA和蛋白质表达的影响。实验选用孵化至11胚龄的鸡胚,在无菌条件下分离其腿肌组织,通过机械法和差速贴壁法分离纯化获得鸡原代成肌细胞。在此基础上,分别在有血清和血清饥饿条件下,在培养液中添加不同浓度的GH (50, 100, 200和500 ng/mL)诱导培养3 h,收获细胞,定量分析IGF1的表达变化。结果表明,体外培养条件下GH可以诱导鸡成肌细胞表达IGF1 mRNA和蛋白质。在相同的GH添加浓度下,IGF1在血清饥饿条件下的表达量高于有血清培养条件。尤其是在200 ng/mL添加浓度下,IGF1表达量极显著高于有血清培养条件。研究结果表明,在血清饥饿培养条件下,200 ng/mL GH能够诱导鸡原代成肌细胞表达IGF1的量达到最高水平。本研究的开展为揭示GH-GHR-IGF1通路调控鸡成肌细胞增殖和分化的机制提供了理论依据,为研究肉鸡骨骼肌发育提供了基本数据和资料。 Endocrine growth hormone (GH) combining with the growth hormone receptor (GHR) to form GH-GHR-IGF1 pathway, played crucial roles in animal growth. Insulin-like growth factor 1 (IGF1), a kind of regulating polypeptide, acted in animal development and physiological regulation. In this study, the effects of different ways and GH concentration on the expression of IGF1 were analyzed in chicken primary myoblast cells. It was benefit for us to determine the appropriate way and GH concentration to induce IGF1 expression, which laid the basis for further analyzing the regulatory mechanism of GH in muscle development. The chicken (Gallus gallus) primary myoblast cells were isolated and cultured by physical operations and differential adhension methods using chicken embryo hatching to 11 d. The different concentrations of GH (50, 100, 200 and 500 ng/mL) were added into the DuIbecco's modified eagIe's medium (DMEM) media with or without fetal bovine (Bos tarus) serum to culture the cells for 3 h. The cells were harvested and the total RNAs were isolated to analyze the expression of IGF1. Results indicated that chicken GH protein could induce the expression of IGF1 both in mRNA and protein levels. Chicken IGF1 were more highly expressed under the condition of serum starvation than that of serum culture at the same GH concentration. The expression of IGF1 reached the highest level when 200 ng/mL GH was added under the condition of serum starvation. It indicated that the best way to induce IGF1 expression was that adding 200 ng/mL GH into DMEM media after chicken primary myoblast cells were serum starvation for 10 h. The results provide the basic data for broiler muscle development and lays the basis for further studying the roles of GH-GHR-IGF1 pathway in myoblast cells proliferation and differentiation.
作者 张丽 王章 李婷 徐海冬 效梅 刘满清 聂庆华 张细权 安立龙 ZHANG Li1, WANG Zhang1, LI Ting1, XU Hai-Dong1 ,XIAO Mei,LIU Man-Qing2,3 ,NIE Qing-Hua2,3, ZHANG Xi-Quan2,3, AN Li-long1( 1 Agricultural College, Guangdong Ocean University, Zhanjiang 524088, China; 2 Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China; 3 College of Animal Science, South China Agricultural University, Guangzhou 510642, China)
出处 《农业生物技术学报》 CAS CSCD 北大核心 2017年第3期511-516,共6页 Journal of Agricultural Biotechnology
基金 国家自然科学基金(Noo.31672412和No.31301958)和农业部鸡遗传育种与繁殖重点实验室开放项目
关键词 鸡成肌细胞 生长激素(GH)蛋白 胰岛素样生长因子1(IGF1)表达 条件优化 Chicken myoblast, Growth hormone(GH) protein, Insulin-like growth factor 1 (IGF1)expression, The appropriate method
作者简介 通讯作者,xqzhang@scau.edu.cn 通讯作者,anlilong@126.com
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