Yang Zisha, Wang Jiajun, Pi Jiang, Hu Di, Xu Junfa, Zhao Yi, Wang Yan
Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, The First Dongguan Affiliated Hospital, Guangdong Medical University, Dongguan, Guangdong, 523713, People's Republic of China.
Institute of Laboratory Medicine, School of Medical Technology, Guangdong Medical University, Dongguan, Guangdong, 523808, People's Republic of China.
J Inflamm Res. 2024 Feb 29;17:1397-1411. doi: 10.2147/JIR.S448372. eCollection 2024.
To investigate the correlation between M1/M2 macrophages (M1/M2 Mφ) and cell death mode under (Mtb) infection.
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).
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.
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.
探讨结核分枝杆菌（Mtb）感染下M1/M2巨噬细胞（M1/M2 Mφ）与细胞死亡模式之间的相关性。
从基因表达综合数据库（GEO）收集原始基因表达谱。从KEGG、FerrDb和GSEA数据库收集与不同细胞死亡模式相关的基因。使用R语言中的limma软件包鉴定基因表达谱中的差异表达基因（DEG）。通过VennDiagram软件包获得M1/M2 Mφ与不同细胞死亡模式的交集基因。通过构建蛋白质-蛋白质相互作用（PPI）网络和受试者工作特征（ROC）曲线分析获得枢纽基因。通过蛋白质免疫印迹法（Western Blot）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枢纽基因的表达。
进行生物信息学分析以筛选Mtb感染的M1 Mφ和不同细胞死亡模式的枢纽基因，分别为NFKB1、TNF、CFLAR、TBK1、IL6、RELA、SOCS1、AIM2；Mtb感染的M2 Mφ和不同细胞死亡模式的枢纽基因，分别为TNF、BIRC3、MAP1LC3C、DEPTOR、UVRAG、SOCS1。结合实验验证，Mtb感染下的M1 Mφ与M2 Mφ相比，死亡（包括凋亡、自噬、铁死亡和焦亡）相关基因表达更高，且NFKB1、TNF、CFLAR、TBK1、IL6、RELA、AIM2、BIRC3、DEPTOR等基因表现出差异表达。
Mtb感染的M1 Mφ中的NFKB1、TNF、CFLAR、TBK1、IL6、RELA、AIM2以及Mtb感染的M2 Mφ中的TNF、BIRC3、DEPTOR可能作为结核病的潜在诊断靶点。在Mtb感染早期，M1 Mφ中的凋亡、自噬、铁死亡和焦亡比M2 Mφ更显著，这可能有助于Mtb感染的Mφ从以M1为主向以M2为主转变，并有助于Mtb的免疫逃逸机制。
