Re-Exploring the Inflammation-Related Core Genes and Modules in Cerebral Ischemia

Mol Neurobiol. 2023 Jun;60(6):3439-3451. doi: 10.1007/s12035-023-03275-1. Epub 2023 Mar 3.

Abstract

The genetic transcription profile of brain ischemic and reperfusion injury remains elusive. To address this, we used an integrative analysis approach including differentially expressed gene (DEG) analysis, weighted-gene co-expression network analysis (WGCNA), and pathway and biological process analysis to analyze data from the microarray studies of nine mice and five rats after middle cerebral artery occlusion (MCAO) and six primary cell transcriptional datasets in the Gene Expression Omnibus (GEO). (1) We identified 58 upregulated DEGs with more than 2-fold increase, and adj. p < 0.05 in mouse datasets. Among them, Atf3, Timp1, Cd14, Lgals3, Hmox1, Ccl2, Emp1, Ch25h, Hspb1, Adamts1, Cd44, Icam1, Anxa2, Rgs1, and Vim showed significant increases in both mouse and rat datasets. (2) Ischemic treatment and reperfusion time were the main confounding factors in gene profile changes, while sampling site and ischemic time were not. (3) WGCNA identified a reperfusion-time irrelevant and inflammation-related module and a reperfusion-time relevant and thrombo-inflammation related module. Astrocytes and microglia were the main contributors of the gene changes in these two modules. (4) Forty-four module core hub genes were identified. We validated the expression of unreported stroke-associated core hubs or human stroke-associated core hubs. Zfp36 mRNA was upregulated in permanent MCAO; Rhoj, Nfkbiz, Ms4a6d, Serpina3n, Adamts-1, Lgals3, and Spp1 mRNAs were upregulated in both transient MCAO and permanent MCAO; and NFKBIZ, ZFP3636, and MAFF proteins, unreported core hubs implicated in negative regulation of inflammation, were upregulated in permanent MCAO, but not in transient MCAO. Collectively, these results expand our knowledge of the genetic profile involved in brain ischemia and reperfusion, highlighting the crucial role of inflammatory disequilibrium in brain ischemia.

Keywords: Brain ischemia; Inflammation; Reperfusion injury; Thrombo-inflammation.

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Brain Ischemia* / metabolism
  • Galectin 3 / metabolism
  • Humans
  • Infarction, Middle Cerebral Artery / genetics
  • Inflammation / genetics
  • Mice
  • RNA, Messenger
  • Rats
  • Reperfusion Injury* / genetics
  • Stroke* / genetics
  • rho GTP-Binding Proteins / metabolism

Substances

  • Galectin 3
  • RNA, Messenger
  • RHOJ protein, human
  • rho GTP-Binding Proteins
  • Nfkbiz protein, mouse
  • Adaptor Proteins, Signal Transducing