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 deposi⁃ tion in goats based on transcriptome sequencing analysis

  • 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(IGFBP7IRX3)。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、AKTMTOR)和脂肪分化标志基因(PPARγ、FASCEBPα)的表达量极显著高于对照组。50 µmol/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、FASCEBPα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。

     

    Abstract: 【Objective】This study was mainly based on transcriptome sequencing(RNA-Seq)to analyze the mechanism of resveratrol promoting intramuscular fat deposition in Nubian goats,in order to find the candidate genes and regulatory 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. Differentially expressed genes(DEGs) screeningFold Change ≥ 2,false discovery rate(FDR) <0.05was 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 differentiation 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 pathways,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 pathway 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 50 μmol/L RSV induced muscle cells,it was found that the lipid droplets were extremely significantly more than those in the control group,and the expression levels of P13K,AKT,MTOR,PPARγ,FAS and CEBPα were extremely significantly higher than those in the control group.【Conclusion】 In summary,based on RNA-Seq analysis of the mechanism of resveratrol promoting intramuscular fat deposition in Nubian goats,combined with its induction of myocyte adipogenic differentiation,it is speculated that resveratrol may promote the deposition of intramuscular fat in Nubian goats through P13K-AKT signaling pathway.

     

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