LncRNA GAS5 ameliorates intestinal barrier injury by targeting the miR-223-3p/FBXW7 axis and inactivating NF-κB signaling in vitro and in vivo

Sepsis is a potentially lethal syndrome that leads to multiple organ dysfunction. LncRNA GAS5 is closely related to sepsis; however, its detailed functions and mechanism in sepsis-triggered intestinal barrier dysfunction are unclear. In this study, NCM460 cells were stimulated with lipopolysaccharide (LPS) to mimic septic intestinal injury in vitro, and a sepsis mouse model was established via the cecum ligation and perforation method. Terminal deoxynucleotidyl transferase dUTP nick end labeling staining was performed for cell Apoptosis evaluation. RNA and protein levels were examined by RT-qPCR and western blotting, respectively. Additionally, cell permeability and intestinal mucosa permeability were measured. ELISA was utilized to detect inflammatory cytokine production. H&E staining was conducted for histologic examination of the intestine. Luciferase reporter and RNA pull-down assays were employed to verify the interaction between GAS5, miR-223-3p, and FBXW7. The results showed that GAS5 was downregulated in LPS-exposed NCM460 cells as well as the intestine of septic mice. GAS5 overexpression mitigated LPS-triggered intestinal epithelial cell damage and Apoptosis in vitro and reduced pathological damage, inflammation, and intestinal hyperpermeability in septic mice. GAS5 upregulated FBXW7 by interacting with miR-223-3p. Depletion of FBXW7 reversed the protective effects of GAS5 overexpression in vitro. Additionally, GAS5 overexpression inactivated NF-κB signaling in LPS-stimulated NCM460 cells. NF-κB inactivation exerted effects in septic mice similar to those observed with GAS5 overexpression. In conclusion, GAS5 ameliorates sepsis-triggered intestinal barrier disruption by mediating the miR-223-3p/FBXW7 axis and inactivating NF-κB signaling. © 2025 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

FBXW7; GAS5; NF‐κB; inflammation; intestinal barrier dysfunction; lipopolysaccharide; sepsis.