【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1μmol/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。更多还原

【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 with1 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-1was 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 upregulated 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.更多还原