Mechanism of Disulfide Death-Driven Intestinal Epithelial Injury in Neonatal Necrotizing Enterocolitis and Exploration of Potential Therapeutic Targets

Background Neonatal necrotizing enterocolitis (NEC) remains a devastating gastrointestinal emergency in premature infants, characterized by abrupt onset and rapid progression to transmural necrosis. Recent evidence suggests that disulfidptosis, a novel form of regulated cell death driven by disulfide stress, may play a pivotal role in various inflammatory diseases. However, its specific contribution to NEC pathogenesis remains largely unexplored. Methods We integrated multi-omic bioinformatic analyses with in vitro experimental validation. Disulfidptosis-related hub genes were identified via weighted gene co-expression network analysis (WGCNA) and differentially expressed gene (DEG) analysis using GEO datasets (GSE46619, GSE297483), with diagnostic efficacy evaluated by ROC curves. In vitro validation was conducted in LPS-stimulated Caco-2 cells, utilizing tris(2-chloroethyl) phosphate (TCEP) to assess disulfidptosis inhibition. Results NEC tissues exhibited significantly elevated disulfidptosis scores, which correlated positively with disease severity. Integrating WGCNA and DEGs identified 147 core genes primarily enriched in inflammatory signaling and intercellular communication. Among these core genes (TKTL1, PFKFB3, SLC2A3, and SLC2A14), TKTL1 exhibited the highest diagnostic accuracy (AUC = 0.892) and were closely associated with altered immune infiltration, supporting a 'metabolic-inflammatory axis' in NEC. In vitro , LPS-stimulated Caco-2 cells manifested definitive disulfidptosis hallmarks—NADP⁺ depletion, cystine accumulation, and F-actin collapse—synchronized with barrier failure. Pharmacological inhibition via TCEP successfully stabilized the cellular redox state, restored cytoskeletal integrity, and attenuated IL-6/TNF-α secretion, thereby preserving epithelial function. Conclusion his study identified disulfidptosis as a critical driver of intestinal epithelial injury in NEC. Targeting disulfidptosis-related pathways may offer a promising diagnostic and therapeutic strategy for neonatal intestinal injury.