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.