Objective: To investigate the effect of Silent information regulator 1 (Sirt1) on the expression profile of long non-coding RNA (lncRNA) in macrophages upon lipopolysaccharide (LPS) treatment. Methods: Peritoneal macrophages (PM) were isolated from nine wild-type C57BL/6 male mice (wild-type group) and nine myeloid-specific Sirt1 knock-out mice (knock-out group). RNA samples were extracted from macrophages stimulated with 1 μg/ml LPS. Sequencing and the differentially expressed lncRNA were screened after the RNA was quantified. The threshold set for up-and down-regulated genes was a fold change (wild-type group/knock-out group) ≥2 and P≤0.05. Afterwards, gene ontology (GO) and pathway enrichment analysis were conducted and co-expression network map was constructed. Results: Four hundred and forty five lncRNA genes were differentially expressed (185 lncRNA genes were up-regulated and 260 lncRNA genes were down-regulated). Two hundred mRNA genes were differentially expressed (113 mRNA genes were up-regulated and 87 mRNA genes were down-regulated). It was found that the differentially expressed lncRNA genes and the predicted corresponding target genes were mainly distributed in the regions of biological processes of macrophage inflammatory response, macrophage chemotaxis and cell metabolism by GO and pathway enrichment analysis. Conclusion: lncRNA expression profile changes significantly in LPS induced macrophages isolated from Sirt1 knock out mice, which is closely related to the function of macrophages.
目的: 探讨沉默信息调节因子1(Sirt1)对脂多糖(LPS)诱导小鼠巨噬细胞长链非编码RNA(lncRNA)表达谱的影响。 方法: 选取野生型C57BL/6雄性小鼠9只(野生型组)和髓系特异性敲除型小鼠9只(敲除型组),分离腹腔巨噬细胞,1 μg/ml的LPS处理细胞,提取总RNA并进行质量检测,合格后进行测序实验,以差异倍数(野生型组/敲除型组)≥2且P≤0.05为纳入标准,检测野生型组和敲除型组中lncRNA差异表达谱,并对差异lncRNA进行基因本体(GO)分析和信号通路分析,构建共表达网络图,从而了解lncRNA参与的生物学功能。 结果: 两组比较,差异表达的lncRNA基因共445个,其中185个基因表达上调,260个基因表达下调。差异表达的mRNA基因共200个,其中113个基因表达上升和87个基因表达下降。通过GO分析和信号通路分析发现差异表达的lncRNA基因及其共表达的mRNA基因主要富集于影响巨噬细胞的炎症反应,巨噬细胞的渗漏,细胞的代谢等生物过程。 结论: Sirt1敲除之后,LPS诱导巨噬细胞中lncRNA表达谱发生明显变化,与巨噬细胞功能的改变密切相关。.
Keywords: Gene expression profiling; Lipopolysaccharides; Macrophages; RNA, untranslated; Silent information regulator proteins.