Nitrate enhances gastric mucosa defence and repair process in ethanol-induced gastric ulcer rats via the Notch-Tff2 pathway

Alcohol consumption is associated with high rates of gastric ulcer incidence and recurrence, with no effective prevention and treatment methods. Nitrates play key roles in systemic disease development. This study aimed to explore nitrate preventive effects on ethanol-induced gastric ulcers. A rat model of ethanol-induced gastric ulcers was established to explore the protective effect of nitrate, and an in vitro model was used to verify the effect of nitrate on the migration of human gastric epithelial cells. Nitrate significantly alleviated ethanol-induced gastric mucosal haemorrhage, oedema, and inflammation, while enhancing mucous layer thickness and secretion of gastric protective factors. Transcriptome analysis and western blot validation identified trefoil peptide factor 2 as a crucial gene and Notch as a vital pathway protein. In rats with knockdown of trefoil peptide factor 2, nitrate protection of the gastric mucosa was almost absent. Furthermore, promotion of wound repair by nitrates diminished in human gastric epithelial cells with a knockout of trefoil peptide factor 2. Nitrate downregulates the Notch pathway, which is excessively activated by ethanol in vivo and in vitro. Notch pathway activation diminishes the nitrate-induced migratory repair capacity of cells. Results from Cut-Tag and dual-luciferase reporter assays indicated that a Notch pathway transcription factor negatively affected trefoil peptide factor 2 expression. Nitrate strengthened gastric mucosal barriers, enhanced mucosal defence, and promoted early epithelial repair in ethanol-induced gastric ulcers via the Notch-trefoil peptide factor 2 pathway. Our study provides useful insights into gastric ulcer prevention in patients who consume alcohol.