Abstract: The purpose of this study was to look into the toxicity of arecoline on human stomach mucosal epithelial cells GES-1. GES-1 cells were treated with different concentrations (60, 90, 120, 150 μg/mL) of arecoline, detection of GES-1 cell viability, reactive oxygen species levels (ROS) and mitochondrial membrane potential (MMP). The levels of glutathione (GSH), superoxide dismutase (SOD), lactate dehydrogenase (LDH), adenosine triphosphate (ATP) enzymes were measured in GES-1 cellls. And the leves of tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), vascular endothelial growth factor (VEGF), transforming growth factor-β (TGF-β) were measured in GES-1 cells. Compared with the control group, the shape of GES-1 cells was altered after arecoline stimulation, with the lowest cell viability rate reduced to 24%, the highest reactive oxygen species level increased to 12.5 fold, and the lowest mitochondrial membrane potential decreased to 10.4%. Under 150 μg/mL arecoline stimulation, T-ATPase and Na⁺K⁺-ATPase activity decreased to 42.31% and 39.84%, GSH and SOD decreased by 40.23% and 34.1%, respectively, LDH increased by 30.46%, TNF-α, IL-6, VEGF and TGF-β levels increased by 4.67%, 10.2%, 23.14% and 22.83%. The results indicate that arecoline can produce oxidative stress and induce inflammatory factors that cause toxic damage to GES-1 cells.