Kaempferol Attenuates Oxidative Stress-Induced Injury in Gastric Mucosal Cells by Activating Nrf2/GPX4 Axis to Inhibit Ferroptosis.

OBJECTIVE: This study explores how Kaempferol (KAE) protects against oxidative stress-induced damage by suppressing ferroptosis via the Nrf2/GPX4 axis in gastric mucosal cells. METHODS: Human gastric epithelial cells (GES-1) were treated with H₂O₂ to induce oxidative damage, following pretreatment with varying doses of KAE. Cell vitality was assessed by the CCK-8 experiment, apoptosis was monitored using flow cytometry, and intracellular ROS levels and lipid peroxidation were determined by fluorescence probes. Intracellular malondialdehyde (MDA), glutathione (GSH/GSSG ratio), and Fe²⁺ were measured using biochemical assays. Expression and cellular distribution of Nrf2, GPX4, SLC7A11, and ACSL4 were assessed using Western blot analysis and immunofluorescence techniques. Additionally, the Nrf2-specific inhibitor ML385 was employed to confirm the role of the Nrf2/GPX4 axis. RESULTS: KAE (0-40 μM) was non-toxic and enhanced GES-1 cell viability under H₂O₂-induced stress, with optimal protection at 10 μM. It reduced ROS, lipid peroxidation, MDA, and Fe²⁺ levels, while increasing the GSH/GSSG ratio. KAE also influenced ferroptosis-associated proteins by increasing GPX4 and SLC7A11 expression while reducing ACSL4 levels. Additionally, it promoted Nrf2 nuclear translocation. These effects were attenuated by the Nrf2 inhibitor ML385, indicating involvement of the Nrf2/GPX4 axis. CONCLUSION: KAE protects against H₂O₂-induced gastric epithelial damage through activating the Nrf2/GPX4 axis, thereby lowering oxidative injury and ferroptotic processes, and offering a potential therapeutic strategy for gastric mucosal protection.