Abstract
BACKGROUND: Intestinal ischemia-reperfusion (II/R) injury is a severe clinical condition in which regulated cell death programs-including pyroptosis and necroptosis-have emerged as key drivers of tissue damage and inflammation. We sought to delineate cell-death-related molecular signatures and candidate therapeutic targets in II/R injury. METHODS: We obtained transcriptome datasets from Gene Expression Omnibus (GEO) databases for mice (GSE96733, GSE232246) and humans (GSE37013). We cross-referenced genes associated with necroptosis and pyroptosis with differentially expressed genes to identify death-related features. Hub genes were identified through the topological structure of protein interaction networks and validated using an internal validation set, an independent validation set, WGCNA, and qRT-PCR. These genes were also associated with immune cell infiltration. Drug-gene interactions were predicted using DGIdb and verified through molecular docking. RESULTS: We identified 1,027 differentially expressed genes (DEGs) in the training set and derived 7 cell death-related differentially expressed genes (DCDEGs) by intersecting gene sets associated with necroptosis and pyroptosis. PPI-based prioritization identified four hub genes-Il1β, Ripk3, Sting1 (Tmem173), and Tnfaip3-suggesting cross-regulatory interactions between inflammation and cell death in ischemia-reperfusion pathology. These hub genes were validated using WGCNA analysis and an internal validation set. Immune infiltration analysis indicated significant correlations between hub genes and multiple immune compartments. A predictive model showed good discrimination in the discovery data, and 54 candidate drugs targeting the hub genes were retrieved. qRT-PCR confirmed dysregulation of three hub genes. CONCLUSION: Il1β, Ripk3, Sting1, and Tnaip3 were identified as hub genes associated with necroptosis and pyroptosis in intestinal ischemia-reperfusion (II/R) injury. This study provides a reproducible framework and identifies testable targets for translational exploration.