Abstract

Background Necrotizing enterocolitis (NEC) is a severe gastrointestinal disease affecting preterm neonates, characterized by intestinal inflammation, epithelial injury, and microbial dysbiosis. Prebiotics such as short-chain fructooligosaccharides (scFOS) have been proposed as nutritional interventions to support intestinal health, but their protective mechanisms in NEC remain incompletely understood. Methods Using a neonatal mouse model of NEC, we investigated the protective effects of scFOS supplementation on intestinal injury, regeneration, inflammation, and microbial composition. Histological analysis, immunostaining, cytokine quantification, quantification of Lactobacillus abundance, Western blotting, and phosphoproteomics were employed to assess intestinal damage and underlying signaling pathways. Results scFOS-treated NEC mice exhibited significantly improved intestinal architecture, reduced histological injury scores, and enhanced epithelial regeneration, as evidenced by restored Ki67 and β-catenin expression. Inflammatory cytokine expression (TNF-α, IL-6) was significantly attenuated, and the abundance of Lactobacillus was increased, indicating a partial correction of NEC-associated dysbiosis. Mechanistically, phosphoproteomic profiling and Western blot analysis revealed that NEC suppressed AKT phosphorylation and activated GSK-3β, leading to β-catenin degradation and impaired repair. scFOS supplementation restored AKT/GSK-3β signaling, promoting mucosal regeneration. Conclusion scFOS protects against NEC-associated intestinal injury through coordinated effects on epithelial regeneration, inflammation, and microbial composition. These benefits are mediated, at least in part, by reactivation of the AKT/GSK-3β/β-catenin signaling axis. These findings support the potential of scFOS as a promising nutritional strategy for NEC prevention in preterm infants.