Identification of key hub genes in spinal cord ischemia-reperfusion injury via integrated bioinformatics analysis and in vivo validation

通过整合生物信息学分析和体内验证,鉴定脊髓缺血再灌注损伤中的关键枢纽基因

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Abstract

Spinal cord ischemia-reperfusion injury (SCII) often causes neurological damage and devastating sensory and motor dysfunction. Identifying key genes and signaling pathways in SCII progression may provide novel therapeutic targets. Two gene expression datasets (GSE138966 and GSE167274) were obtained from the Gene Expression Omnibus database. Differentially expressed genes were identified using R software, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Hub genes were screened via Venn analysis, and a protein-protein interaction (PPI) network was constructed using Cytoscape software. Key hub genes were validated by qRT-PCR in a rat SCII model. A total of 99 hub genes were identified, including 60 up-regulated and 39 down-regulated genes. KEGG analysis revealed significant enrichment in MAPK, cAMP, and Rap1 signaling pathways. PPI network analysis highlighted Ccl2, Mmp9, Itgb1, Timp1, Myd88, and Lgals3 as central nodes. qRT-PCR validation showed persistent up-regulation of Tnc, Thbs2, and S100a10 at 1 h, 24 h, and 48 h post-SCII; early up-regulation of Msn, Lcp1, Lcn2, and Akap12 at 1 h; and delayed up-regulation of Itga5 at 48 h (P < 0.05). This study identifies novel, key SCII-related genes that have been largely overlooked and, for the first time, defines their time-dependent expression patterns via in vivo experimental validation. Our findings provide crucial mechanistic insights and nominate promising therapeutic targets for SCII.

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