Molecular docking results.

BackgroundIntestinal 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.MethodsWe 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.ResultsWe 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.ConclusionIl1β, 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.

背景：肠缺血再灌注（intestinal ischemia–reperfusion, II/R）损伤是一种严重的临床病症，其中包括焦亡（pyroptosis）、坏死性凋亡（necroptosis）在内的程序性细胞死亡程序已成为组织损伤与炎症的核心驱动因素。本研究旨在阐明肠缺血再灌注损伤中与细胞死亡相关的分子特征及潜在治疗靶点。
方法：本研究从基因表达综合数据库（Gene Expression Omnibus, GEO）获取小鼠（GSE96733、GSE232246）及人类（GSE37013）的转录组数据集。将与坏死性凋亡、焦亡相关的基因与差异表达基因进行交叉比对，以筛选细胞死亡相关特征。通过蛋白质相互作用网络的拓扑结构筛选枢纽基因，并采用内部验证集、独立验证集、加权基因共表达网络分析（Weighted Gene Co-expression Network Analysis, WGCNA）及实时荧光定量PCR（quantitative real-time polymerase chain reaction, qRT-PCR）进行验证。同时分析这些枢纽基因与免疫细胞浸润的相关性。利用药物基因相互作用数据库（Drug-Gene Interaction Database, DGIdb）预测药物-基因相互作用，并通过分子对接进行验证。
结果：本研究在训练集中筛选得到1027个差异表达基因（differentially expressed genes, DEGs），通过与坏死性凋亡、焦亡相关基因集取交集，获得7个细胞死亡相关差异表达基因（cell death-related differentially expressed genes, DCDEGs）。基于蛋白质相互作用（protein-protein interaction, PPI）网络的优先级分析筛选出4个枢纽基因：Il1β、Ripk3、Sting1（Tmem173）及Tnfaip3，提示缺血再灌注病理过程中炎症与细胞死亡存在交叉调控相互作用。上述枢纽基因经加权基因共表达网络分析及内部验证集验证有效。免疫浸润分析显示，枢纽基因与多个免疫细胞亚群存在显著相关性。预测模型在发现数据集上表现出良好的区分能力，共检索到54种靶向该类枢纽基因的候选药物。实时荧光定量PCR验证证实3个枢纽基因表达失调。
结论：本研究鉴定出Il1β、Ripk3、Sting1及Tnaip3为肠缺血再灌注（II/R）损伤中与坏死性凋亡、焦亡相关的枢纽基因。本研究构建了可重复的分析框架，并为转化研究确定了可验证的治疗靶点。