DS0384 Alleviates Necrotizing Enterocolitis: Secretes N-carbamyl glutamic Acid and Participates in Lipid Metabolism and Lipid Peroxidation Processes.

Abstract

Necrotizing enterocolitis (NEC) is a life-threatening inflammatory bowel disease linked to gut microbiome dysbiosis. This study evaluates the efficacy of Limosilactobacillus reuteri strain DS0384, which secretes N-carbamyl glutamic acid (NCG), in modulating lipid peroxidation and inflammatory pathways in NEC. After pretreatment with DS0384, NEC mouse model was induced by gavage with bacteria-containing formula. NCG levels in the ileum were measured via CE-TOFMS metabolomic analysis. Additionally, rat small intestinal epithelial IEC-6 cells were exposed to lipopolysaccharide (LPS), treated with DS0384 DNA (D-DNA), and/or transfected to overexpress fatty acid synthase (FASN) and Toll-like receptor 4 (TLR4). Lipid peroxidation, peroxidation and inflammatory factors and NF-κB pathways were analysed. Immunofluorescence was used to measure the expression levels of ZO-1 and TLR4 in the ileum. DS0384 treatment significantly reduced more histological abnormalities, apoptosis, and TLR4 expression in NEC mice, while restoring NCG levels, downregulating FASN and inhibiting lipid peroxidation and inflammation. Pre-treatment with D-DNA maintained cell vitality, reduced apoptosis, and suppressed TLR4/NF-κB-mediated inflammasome activation. Overexpression of FASN or TLR4 reversed these effects. DS0384 is a promising therapeutic against NEC, enhancing gut barrier integrity and modulating inflammatory and oxidative responses, suggesting potential clinical benefits in preventing NEC progression.