动物营养学报 >

2021 , Vol. 33 >Issue 4: 2263 - 2277

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.04.045

分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

基于转录组测序的油酸诱导延边牛骨骼肌卫星细胞成脂分化差异表达基因的分析

  • 孙建富 ,
  • 孙斌 ,
  • 张军芳 ,
  • 王英 ,
  • 崔岩 ,
  • 李强 ,
  • CHOI Seong H ,
  • SHIN Jong S ,
  • 李香子
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  • 1. 延边大学东北寒区肉牛科技创新教育部工程研究中心, 延吉 133002;
    2. 韩国忠北国立大学动物生命科学学院, 清州361-763;
    3. 韩国江原国立大学校动物生命科学学院, 春川200-701
孙建富(1980-),男,吉林磐石人,博士研究生,从事肉牛脂肪代谢与功能细胞研究。E-mail:sunjianfu4379@163.com

收稿日期: 2020-10-22

  网络出版日期: 2021-04-15

基金资助

国家自然科学基金——PLIN2在油酸诱延边黄牛骨骼肌细胞向脂肪细胞分化的作用及其机制研究（31860653）；东北寒区肉牛科技创新教育工程研究中心项目；高等学校学科创新引智计划（D20034）

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Analysis of Differentially Expressed Genes for Adipogenic Differentiation of Yanbian Bovine Skeletal Muscle Satellite Cells Induced by Oleic Acid Based on Transcriptome Sequencing

  • SUN Jianfu ,
  • SUN Bin ,
  • ZHANG Junfang ,
  • WANG Ying ,
  • CUI Yan ,
  • LI Qiang ,
  • CHOI Seong ,
  • SHIN Jong ,
  • LI Xiangzi
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  • 1. Northeast Cold Region Beef Cattle Science and Technology Innovation Ministry of Education Engineering Research Center, Yanbian University, Yanji 133002, China;
    2. College of Animal Life Sciences, Chungbuk National University, Cheongju 361-763, South Korea;
    3. College of Animal Life Sciences, Gangwon National University, Chuncheon 200-701, South Korea

Received date: 2020-10-22

  Online published: 2021-04-15

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摘要

本研究旨在研究油酸（OA）引起延边牛骨骼肌卫星细胞成脂分化途径的分子调控机制。对从12日龄延边牛中分离得到的骨骼肌卫星细胞进行体外培养，加入不同浓度OA的诱导分化培养液，进行分化处理，试验设1个空白对照组[CON组，5%马血清（HS）]，3个不同浓度OA诱导组：OAL组（5% HS+50 μmol/L OA）、OAM组（5% HS+100 μmol/L OA）、OAH组（5% HS+200 μmol/L OA），诱导96 h后，对延边牛骨骼肌卫星细胞进行了转录组测序及分析。结果表明：共得到13 951条单基因，对4个组差异表达基因进行比较发现，与CON组相比，OAL组有3 412个差异表达基因，OAM组有1 045个差异表达基因，OAH组有1 428个差异表达基因，各组之间共有278个差异表达基因。GO富集分析显示，差异基因参与了多种生物过程，包括代谢过程、细胞过程、生物调节过程等；在分子功能类别上，大多数的基因功能与结合活性、转录调节活性、催化活性等相关；在细胞组分范畴内，大部分的基因被富集到细胞、细胞器、膜等。KEGG富集分析表明，差异表达基因主要富集通路为单磷酸腺苷激活的蛋白激酶（AMPK）信号通路、过氧化物酶体增殖物激活受体（PPAR）信号通路、脂肪酸代谢通路、脂肪酸降解通路等。不同浓度OA差异基因荧光定量PCR结果显示，所选的5个差异基因与转录组测序结果基本一致，表明了测序结果的可靠性。本试验完成了OA诱导延边牛骨骼肌卫星细胞成脂分化的转录组测序分析，获取差异基因的功能注释信息，初步揭示了OA诱导牛骨骼肌卫星细胞成脂分化的潜在基因和通路。

关键词： 转录组测序; 油酸; 延边牛骨骼肌卫星细胞; 成脂分化

本文引用格式

孙建富 , 孙斌 , 张军芳 , 王英 , 崔岩 , 李强 , CHOI Seong H , SHIN Jong S , 李香子 . 基于转录组测序的油酸诱导延边牛骨骼肌卫星细胞成脂分化差异表达基因的分析[J]. 动物营养学报, 2021 , 33(4) : 2263 -2277 . DOI: 10.3969/j.issn.1006-267x.2021.04.045

Abstract

This study aimed to study the molecular regulation mechanism of oleic acid (OA)-induced adipogenic differentiation of Yanbian bovine skeletal muscle satellite cells. The skeletal muscle satellite cells isolated from the 12-day-old Yanbian cattle were cultured in vitro, and different OA concentrations were added to induce differentiation media for differentiation. The experiment set up a blank control group[CON group, 5% horse serum (HS)], 3 OA induction groups:OAL group (5% HS+50 μmol/L), OAM group (5% HS+100 μmol/L), OAH group (5% HS+200 μmol/L). After 96 h of induction, transcriptome sequencing and analysis of Yanbian bovine skeletal muscle satellite cells were performed. The results showed as follows:a total of 13 951 single genes were obtained. The differentially expressed genes of the 4 groups were compared and found that compared with the CON group, the OAL group had 3 412 differentially expressed genes, the OAM group had 1 045 differentially expressed genes, and there were 1 428 differentially expressed genes in OAH group, and there were a total of 278 differentially expressed genes between each groups. GO enrichment analysis showd that differential genes were involved in a variety of biological processes, including metabolic processes, cellular processes, and biological regulation processes. In terms of molecular functions, most gene functions were related to binding activity, transcription regulation activity, and catalytic activity; in the category of cell components, most of the genes were enriched in cells, organelles, membranes and so on. KEGG enrichment analysis showed that the main enrichment pathways of differentially expressed genes were AMP-activated protein kinase signaling pathway, peroxisome proliferators-activated receptors signaling pathway, fatty acid metabolism pathway, fatty acid degradation pathway, etc. The results of real-time PCR showed that the five genes were consistent with the transcriptome sequencing results, which indicated the reliability of the sequencing results. This experiment completes the transcriptome sequencing analysis of the adipogenic differentiation of Yanbian bovine skeletal muscle satellite cells induced by OA, obtained functional annotation information of different genes, and initially revealed the potential genes and pathways of OA-induced adipogenic differentiation of bovine skeletal muscle satellite cells. In-depth exploration of the molecular mechanism of OA-induced adipogenic differentiation of bovine skeletal muscle satellite cells and related metabolic pathways lay the foundation.

Key words： transcriptome sequencing; oleic acid; Yanbian bovine skeletal muscle satellite cells; adipogenic differentiation

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