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作 者:郭莉 张燕[1,2,3] 罗文萍 赵天宇[1,2,3] 杨德琴 GUO Li;ZHANG Yan;LUO Wen-ping;ZHAO Tian-yu;YANG De-qin(Stomatological Hospital of Chongqing Medical University,Chongqing 401147,China;Chongqing Municipal Key Laboratory of Oral Diseases and Biomedical Sciences,Chongqing 401147,China;Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education Institutions,Chongqing 401147,China)
机构地区:[1]重庆医科大学附属口腔医院,重庆401147 [2]口腔疾病与生物医学重庆市重点实验室,重庆401147 [3]重庆市高校市级口腔生物医学工程重点实验室,重庆401147
出 处:《四川大学学报(医学版)》2022年第3期444-451,共8页Journal of Sichuan University(Medical Sciences)
基 金:国家自然科学基金面上项目(No.319770783);重庆市医学重点学科建设项目(2011年,No.55,《牙体牙髓病学》);重庆市教育委员会重点项目(No.KJZD-K201900402)资助。
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。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 with1μmol/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-1βwas 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-1βexpression 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-1βby 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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