The effects of ethanol on gastric acid secretion remain controversial. The present study examines the effect of low-dose (2%) short term (15-20min) ethanol exposure on gastric acid secretion via a potential interaction with AMP-activated protein kinase (AMPK). Real-time fluorescence digital imaging was used to provide functional evidence for the interaction of ethanol and AMPK in modulating secretagogueinduced acid secretion. Individual rat gastric glands were loaded with the pH-sensitive dye BCECF and the secretagogues carbachol (200 µM) or histamine (200 µM) were added to induce secretion. Rates of pH recovery were calculated as ΔpH i /Δt. In one series of experiments, secretagogue-induced acid secretion was inhibited by 2% ethanol, or the AMPK activator AICAR monophosphate (AICAR) (20 mM). In a separate series, 2% ethanol was added in combination with compound C (20 µM), an AMPK inhibitor, to prevent activation of AMPK. 2% ethanol significantly suppressed stimulated acid secretion.In order to confirm modulation of AMPK activity by ethanol, the specific AMPK inhibitor compound C was used, which reversed the inhibitory effects of ethanol on stimulated acid secretion. This study demonstrates that low dose ethanol (2%) inhibits secretagoguedependent acid secretion by activation of the AMPK pathway in rat gastric parietal cells.