■研究结核分枝杆菌(Mtb)感染下M1/M2巨噬细胞(M1/M2Mφ)与细胞死亡模式的相关性。
■从基因表达综合(GEO)数据库收集原始基因表达谱。从KEGG收集与不同细胞死亡模式相关的基因,FerrDb和GSEA数据库。使用R中的limma包鉴定了基因表达谱的差异表达基因(DEGs)。通过VennDigraph包获得具有不同细胞死亡模式的M1/M2Mφ的相交基因。通过构建蛋白质-蛋白质相互作用(PPI)网络和接受者工作特征(ROC)曲线分析获得Hub基因。通过Western印迹和定量实时聚合酶链反应(qRT-PCR)验证细胞死亡模式标记基因和Hub基因的表达。
■进行了生物信息学分析,以筛选Mtb感染的M1Mφ的Hub基因和不同的细胞死亡模式,命名NFKB1,TNF,CFLAR,TBK1,IL6,RELA,SOCS1,AIM2;Mtb感染的M2Mφ的Hub基因和不同的细胞死亡模式,命名TNF,BIRC3,MAP1LC3C,DEPTOR,UVRAG,SOCS1.结合实验验证,Mtb感染下M1Mφ表现出较高的死亡表达(包括凋亡、自噬,铁性凋亡,和焦亡)基因与M2Mφ和NFKB1,TNF,CFLAR,TBK1,IL6,RELA,AIM2、BIRC3、DEPTOR显示差别表达。
■NFKB1,TNF,CFLAR,TBK1,IL6,RELA,Mtb感染M1Mφ中的AIM2,和TNF,Mtb感染的M2Mφ中的BIRC3、DEPTOR可能作为结核病的潜在诊断目标。在Mtb感染的早期阶段,凋亡,自噬,铁性凋亡,M1Mφ的焦化发生比M2Mφ更显著,这可能有助于Mtb感染的Mφ从M1显性转变为M2显性,并有助于Mtb的免疫逃逸机制。
UNASSIGNED: To investigate the correlation between M1/M2 macrophages (M1/M2 Mφ) and cell death mode under Mycobacterium tuberculosis (Mtb) infection.
UNASSIGNED: Raw gene expression profiles were collected from the Gene Expression Omnibus (GEO) database. Genes related to different cell death modes were collected from the KEGG, FerrDb and GSEA databases. The differentially expressed genes (DEGs) of the gene expression profiles were identified using the limma package in R. The intersection genes of M1/M2 Mφ with different cell death modes were obtained by the VennDiagram package. Hub genes were obtained by constructing the protein-protein interactions (PPI) network and Receiver Operating Characteristic (ROC) curve analysis. The expression of cell death modes marker genes and Hub genes were verified by Western Blot and Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR).
UNASSIGNED: Bioinformatics analysis was performed to screen Hub genes of Mtb-infected M1 Mφ and different cell death modes, naming NFKB1, TNF, CFLAR, TBK1, IL6, RELA, SOCS1, AIM2; Hub genes of Mtb-infected M2 Mφ and different cell death modes, naming TNF, BIRC3, MAP1LC3C, DEPTOR, UVRAG, SOCS1. Combined with experimental validation, M1 Mφ under Mtb infection showed higher expression of death (including apoptosis, autophagy, ferroptosis, and pyroptosis) genes compared to M2 Mφ and genes such as NFKB1, TNF, CFLAR, TBK1, IL6, RELA, AIM2, BIRC3, DEPTOR show differential expression.
UNASSIGNED: NFKB1, TNF, CFLAR, TBK1, IL6, RELA, AIM2 in Mtb-infected M1 Mφ, and TNF, BIRC3, DEPTOR in Mtb-infected M2 Mφ might be used as potential diagnostic targets for TB. At early stage of Mtb infection, apoptosis, autophagy, ferroptosis, and pyroptosis occurred more significantly in M1 Mφ than that in M2 Mφ, which may contribute to the transition of Mtb-infected Mφ from M1-dominant to M2-dominant and contribute to the immune escape mechanisms of Mtb.