Acrylamide (ACR) is a food toxicant detected in over-thermal cooked foods, especially carbohydrate-rich foodstuff. ACR induced toxicity in different organs in the body. The current study was planned to evaluate the possible ameliorative effect of Thymoquinone (TQ) against ACR-induced hepatotoxicity. Twenty-four adult male Wistar albino rats were segregated into four groups (seven rats/group): Control group (saline once daily), TQ group (20 mg/kg once daily), ACR group (20 mg/kg once daily), ACR+TQ group (20 mg/kg TQ plus 20 mg/kg ACR once daily). All treatment of the study via oral gavage for 28 days. Specimens of serum and hepatic tissue were taken on the last day of the experiment. Biochemical parameters include aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), cholesterol (CHO), triglycerides (TG), low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C) were estimated. Furthermore, oxidative cascade markers including malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and reduced glutathione (GSH) were evaluated. Histopathological investigation of the liver tissue was performed. ACR caused liver damage indicated by reducing the activity of liver enzymes and loss of the hepatic antioxidant activity of CAT, SOD, and GPx, and GSH content with a significant elevation in MDA level in serum. Besides, ACR induced histopathological alterations in the hepatic tissue. However, TQ alleviated the hepatic damage, oxidative stress, and histopathological alteration of liver tissue induced by ACR. Overall, these results concluded that TQ improved ACR-induced liver damage by its antioxidant mechanism.