Formononetin attenuates ethanol-induced gastric ulcer injury by targeting ALDH2 to inhibit oxidative stress.

Ethanol-induced gastric ulceration is primarily driven by oxidative stress, mitochondrial dysfunction, and gastric epithelial cell injury. Although formononetin (Fmn), a naturally occurring isoflavone, possesses well-documented antioxidant and anti-inflammatory properties, its protective role and underlying molecular mechanisms in ethanol-induced gastric injury remain unclear. In this study, ethanol-treated GES-1 gastric epithelial cells were used as an in vitro model to investigate the cytoprotective effects of Fmn. Cell proliferation, apoptosis, oxidative stress, and mitochondrial function were evaluated using biochemical and cellular assays. Potential molecular targets were identified through SwissTargetPrediction and molecular docking, followed by experimental validation, including ALDH2 knockdown, qRT-PCR, and enzymatic activity assays. Fmn significantly attenuated ethanol-induced cellular injury by restoring cell proliferation, suppressing apoptosis, reducing intracellular and mitochondrial reactive oxygen species, decreasing malondialdehyde levels, and enhancing superoxide dismutase activity. Molecular docking demonstrated a strong binding affinity between Fmn and aldehyde dehydrogenase 2 (ALDH2), which was further confirmed by increased ALDH2 mRNA and protein expression as well as enhanced enzymatic activity following Fmn treatment. Functional studies revealed that ALDH2 upregulation plays a pivotal role in preserving mitochondrial integrity and limiting oxidative damage. Collectively, these findings demonstrate that Fmn protects gastric epithelial cells against ethanol-induced injury through ALDH2-mediated suppression of oxidative stress and mitochondrial dysfunction, highlighting its potential as a promising therapeutic candidate for the prevention and treatment of alcohol-induced gastric ulceration.