肾上腺素对民猪脂肪细胞基因表达模式的影响

Effects of adrenaline on gene expression patterns in the adipocytes of Min pigs

为了探究冷刺激对民猪脂肪细胞代谢的影响,试验以体外培养的前脂肪细胞为试验材料,通过添加终浓度为1 000 nmol/L的肾上腺素模拟冷刺激环境,同时设置未添加肾上腺素的对照组,诱导分化7 d后收集细胞进行转录组测序(RNA-seq)分析,筛选出表达水平发生变化的差异基因,对差异表达基因进行GO功能注释和KEGG信号通路富集分析,最后通过qRT-PCR验证试验检测RNA-seq结果的准确性。结果表明:肾上腺素会改变脂肪细胞中基因的表达模式,引起128个基因显著上调,93个基因显著下调,其中游离脂肪酸受体4(FFAR4)基因上调倍数最大,为4.63倍。对差异表达基因进行GO功能注释,在分子功能注释分析中有8个条目显著富集,在生物过程注释分析中有128个条目显著富集,在细胞组分注释分析中无显著富集的条目。对差异基因进行KEGG信号通路富集分析,仅神经活性配体-受体相互作用通路被富集。说明肾上腺素可以改变脂肪细胞原有的代谢模式,与脂肪酸代谢和糖原合成等相关的基因发生了显著变化,同时细胞内外的信号转导也发生了改变。

In order to investigate the effects of cold stress on the metabolism of Min pig adipocytes,preadipocytes cultured in vitro were used as experimental materials,the cold stress environment was simulated by adding a final concentration of 1000 nmol/L adrenaline.At the same time,a control group without adrenaline was set up.After induce differentiation for 7 days,cells were collected for RNA-seq analysis,and differentially expressed genes were screened out.Subsequently,GO function annotation and KEGG signaling pathway enrichment analysis were performed for differentially expressed genes.Finally,the accuracy of RNA-seq results was verified through qRT-PCR experiments.The results showed that adrenaline could alter the expression pattern of genes in adipocytes,causing a significant upregulation of 128 genes and a significant downregulation of 93 genes,in which free fatty acid receptor 4(FFAR4)gene was the most up-regulated with 4.63 fold change.GO functional annotation was used to analyze differentially expressed genes.In the molecular functional annotation analysis,8 terms were significantly enriched in the molecular function ontology.In the biological annotation analysis,128 terms were significantly enriched.In the cell component annotation analysis,there were no significantly enriched terms.KEGG pathway analysis of differential genes showed that only the neuroactive ligand-receptor interaction pathway was enriched.The results showed that adrenaline supplementation could change the original metabolic mode of preadipocytes;the genes related to fatty acid metabolism and glycogen synthesis had significant changes,as well as the signal transduction inside and outside the cells.