全反式维甲酸对巨噬细胞中白介素-1β表达的调控及机制研究
Regulatory Effect of All-Trans Retinoic Acid on the Expression of IL-1β in Macrophages and the Mechanisms Involved
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摘要:目的 探究全反式维甲酸(all-trans retinoic acid, ATRA)对巨噬细胞中白介素(interleukin, IL)-1β表达的调控及作用机制。方法 巨噬细胞经1 μmol/L ATRA处理24 h后转录组测序,筛选差异表达基因,进行KEGG通路分析、GO功能分析和PPI网络分析。不同剂量ATRA处理巨噬细胞24 h后,qRT-PCR和Western blot验证炎症因子IL-1β的表达水平。Western blot和免疫荧光染色检测核因子(neuclear factor, NF)-κB信号和半胱天冬酶-1(caspase-1)的变化。结果 测序结果显示,与空白对照组相比,巨噬细胞经ATRA处理后71个差异表达基因上调,KEGG分析显示上调基因参与IL-17信号通路、肿瘤坏死因子(tumor necrosis factor, TNF)信号通路等,GO分析显示上调基因参与IL-1β的产生、对脂多糖的反应等生物学过程,PPI分析揭示炎症因子,黏附分子和趋化因子为ATRA作用的核心基因。体外实验表明,ATRA呈浓度依赖性促进巨噬细胞中IL-1β的表达,ATRA组中磷酸化(p)-NF-κB、NF-κB和caspase-1的表达较对照组升高(P<0.05),且p-NF-κB发生了核转移。结论 ATRA可能通过激活巨噬细胞中的NF-κB信号和caspase-1促进炎症因子IL-1β的表达。Abstract:Objective To investigate the regulatory effect of all-trans retinoic acid (ATRA) on the expression interleukin-1β (IL-1β) in macrophages and the mechanisms involved.Methods Macrophages were treated with 1 μmol/L ATRA for 24 h before RNA-Sequence. Differentially expressed genes (DEGs) were screened out and analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, gene ontology (GO) functional analysis, and protein-protein interaction networks (PPI) analysis. After treatment with different doses of ATRA for 24 h, the expression of IL-1β was examined with qRT-PCR and Western blot. The activation of NF-κB signaling and caspase-1 was observed by Western blot and immunofluorescence staining.Results Compared with the blank control group, a total of 71 DEGs of macrophages were upregulated in the ATRA treatment group. KEGG analysis showed that the up-regulated DEGs were involved in IL-17 signaling pathway, tumor necrosis factor (TNF) signaling pathway, etc. GO analysis indicated that the up-regulated DEGs were involved in the biological processes of the production of IL-1β, response to lipopolysaccharide, etc. PPI analysis revealed that inflammatory cytokines, adhesion molecules, and chemokines were the key genes that ATRA acted on. In vitro experiments showed that ATRA promoted IL-1β expression in macrophages in a concentration-dependent manner. The expression of p-NF-κB, NF-κB, and caspase-1 were significantly increased by ATRA compared with those of the control group (P<0.05), and p-NF-κB translocated to the cell nucleus in the ATRA group.Conclusion ATRA may promote the expression of IL-1β by activating NF-κB signaling and caspase-1 in macrophages, this study may provide evidence for the immune regulatory function of ATRA on macrophages.
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