基于转录组测序分析白藜芦醇促进山羊肌内脂肪沉积的机制  

Mechanism of resveratrol promoting intramuscular fat deposition in goats based on transcriptome sequencing analysis

作　　者：宋颖 张叁保 徐健健 邹剑伟 连子童 邹菊红 黄艳娜[1] 蒋钦杨[1] SONG Ying;ZHANG San-bao;XU Jian-jian;ZOU Jian-wei;LIAN Zi-tong;ZOU Ju-hong;HUANG Yan-na;JIANG Qin-yang(College of Animal Science and Technology,Guangxi University,Nanning,Guangxi 530004,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530004

出　　处：《南方农业学报》2023年第3期681-690,共10页Journal of Southern Agriculture

基　　金：国家重点研发计划项目(2021YFD1100100);国家现代农业产业技术体系广西创新团队建设项目(nycytxgxcxtd-2021-09)。

摘　　要：【目的】基于转录组测序(RNA-Seq)分析白藜芦醇促进努比亚山羊肌内脂肪沉积的机制,以期找到白藜芦醇影响肌内脂肪沉积的候选基因及调控网络。【方法】采用转录组测序技术对饲料中添加600 mg/kg白藜芦醇组山羊的背最长肌和对照组山羊的背最长肌进行分析。利用edgeR软件进行差异基因(Differentially expressed genes,DEGs)筛选[Fold Change≥2,错误发现率(FDR)<0.05],并对DEGs进行GO功能注释和KEGG富集分析。实时荧光定量PCR(qRT-PCR)检测P13K-AKT信号通路关键基因及脂肪分化标志基因的表达。【结果】600 mg/kg白藜芦醇组山羊肌内脂肪含量极显著高于对照组(P<0.01,下同)。RNA-Seq结果显示共筛选出399个DEGs,包括328个上调基因及71个下调基因。DEGs共注释到48个条目,主要包括细胞过程、单有机体过程、细胞、绑定和催化活性。KEGG通路富集分析发现,DEGs主要富集到252个通路,主要包括P13K-AKT信号通路、MAPK信号通路、AMPK信号通路等与脂肪代谢相关的通路,从中进一步筛选出可能参与肌内脂肪沉积的基因(IGFBP7和IRX3)。qRT-PCR结果显示,试验组山羊背最长肌P13K-AKT信号通路关键基因(P13K、AKT和MTOR)和脂肪分化标志基因(PPARγ、FAS和CEBPα)的表达量极显著高于对照组。50µmol/L RSV诱导肌细胞8 d后发现,脂滴极显著多于对照组,P13K、AKT、MTOR、PPARγ、FAS和CEBPα的表达量极显著高于对照组。【结论】基于RNA-Seq分析白藜芦醇促进努比亚山羊肌内脂肪沉积的机制,结合其诱导肌细胞成脂分化,猜测白藜芦醇可能通过P13K-AKT信号通路促进努比亚山羊肌内脂肪的沉积。【Objective】This study was mainly based on transcriptome sequencing(RNA-Seq)to analyze the mecha-nism of resveratrol promoting intramuscular fat deposition in Nubian goats,in order to find the candidate genes and regu-latory networks of resveratrol affecting intramuscular fat deposition.【Method】The longissimus dorsi muscle of goats in the 600 mg/kg resveratrol group and the control group were analyzed by transcriptome sequencing technology.Differen-tially expressed genes(DEGs)screening[Fold Change≥2,false discovery rate(FDR)<0.05]was performed using edgeR software,and GO function annotation and KEGG enrichment analysis were performed on DEGs.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression of P13K-AKT signaling pathway key genes and fat differen-tiation marker genes.【Result】The results showed that the intramuscular fat content of goats in the 600 mg/kg resveratrol group was extremely significantly higher than that in the control group(P<0.01,the same below).RNA-Seq results showed that 399 differential genes were screened,including 328 up-regulated genes and 71 down-regulated genes.DEGs were annotated to a total of 48 entries,mainly including cellular processes,single-organism processes,cellular,binding,and catalytic activities.KEGG pathway enrichment analysis found that the DEGs were mainly enriched to 252 path-ways,mainly including P13K-AKT signaling pathway,MAPK signaling pathway,AMPK signaling pathway and other pathways related to fat metabolism,from which genes that might be involved in intramuscular fat deposition(IGFBP7,IRX3)were further screened out.The results of qRT-PCR showed that the expression levels of P13K-AKT signaling path-way key genes(P13K,AKT,MTOR)and fat differentiation marker genes(PPARγ,FAS,CEBPα)in longissimus dorsi muscle of goats in the experimental group were extremely significantly higher than those in the control group.After 8 d of 50μmol/L RSV induced muscle cells,it was found that the lipid droplets were extremely significantly more th

关 键 词：转录组测序 白藜芦醇 山羊 肌内脂肪 P13K-AKT 

分 类 号：S827[农业科学—畜牧学]