Acrylamide is one of the most widely used in the industry, which found in potatoes, breads, chips, biscuits and many carbohydrate-rich foods and processed at high temperatures. Therefore, human exposure to this compound is high. The toxicity of this compound has been proved in various studies. Acrylamide also causes genetic, neurological, and cancerous effects. The main toxicity mechanism of acrylamide is oxidative stress. Using a diet containing fruits and vegetables that contain natural antioxidant, they are recommended as the main protective strategy against oxidative stress and liver damage. In this study, the protective effect of gallic acid (10,20,50,100 µM) against oxidative damage induced by acrylamide in hepatocytes isolated from rats was studied. Hepatocytes were prepared from rat liver by collagenase perfusion and then the protective effect of gallic acid on acrylamide-induced cellular toxicity by measurement cell death, ROS production, lipid peroxidation, mitochondrial membrane potential loss and lysosomal membrane damage were investigated. To determine the toxicity of acrylamide, EC50,2h concentration of acrylamide (2 mM concentration) was used acrylamide in EC50,2h concentration caused ROS production rise within 3 hours of incubation, Extension of lipid peroxidation, mitochondrial membrane potential loss and cell death compared to control group (p <0/05). Our results showed that gallic acid at concentrations of 50 and 100 µM can reduce the production of reactive oxygen species, lipid peroxidation, mitochondrial damage, and acrylamide-induced cell death.