A novel triazole derivative ameliorates ethanol-induced gastric ulcer via a NOS2-centered inhibition of the AGE-RAGE pathway.

Ethanol-induced gastric ulcer, driven by intricate inflammatory and oxidative stress pathways, lacks optimally effective treatments. Utilizing an integrated approach combining network pharmacology and experimental validation in vivo using a mouse model and in vitro with gastric epithelial cells, this study elucidates the gastroprotective mechanism of the compound MPTA. Network pharmacology identified NOS2 as a central hub gene, guiding subsequent experimental investigation. In an ethanol-induced ulcer mouse model, MPTA administration dose-dependently ameliorated gastric mucosal damage, suppressed pro-inflammatory cytokines including TNF-α, IL-6, IL-1β and IL-8, elevated TGF-β and NO levels, and reduced oxidative stress; these protective effects were attenuated by a NOS2 inhibitor. Proteomic and molecular analyses demonstrated that MPTA downregulated the AGE-RAGE/NF-κB/p38 MAPK inflammatory pathway and activated the NRF2/HO-1/SOD2 antioxidant axis. In vitro, MPTA enhanced cell viability and migration while diminishing apoptosis and ROS accumulation in gastric epithelial cells, primarily through NOS2-centered regulation of the AGE-RAGE signaling pathway. We conclude that MPTA protects against ethanol-induced gastric injury by inhibiting NOS2 to mitigate inflammatory and oxidative stress via the AGE-RAGE pathway. This study unveils a novel gastroprotective mechanism centered on the downregulation of NOS2, underscoring its potential as a protective candidate for ethanol-induced gastric ulcer.