The goal of this study was to see how thymoquinone (TQ) altered acrylamide (ACR) induced toxicity in rats. The current study was carried out on 28 male Wistar rats divided into 4 experimental groups: control (saline, once daily, PO); TQ group (20 mg/kg, once daily, PO); ACR group (20 mg/kg, once daily, PO). The ACR+TQ group received ACR, TQ as mentioned. Saline, TQ and/or ACR, were administered for 28 days. The serum was separated, and the kidney tissues were collected, in order to assess biochemical and oxidative stress markers, as well as histological and immunohistochemical alterations. Levels of creatinine and urea were increased significantly in ACR-treated groups when compared to control. The concentration of malondialdehyde (MDA) in the kidney tissues of the ACR-treated groups were notably substantially higher than in the other groups. Catalase (CAT), Superoxide dismutase (SOD), Glutathione peroxidase (GSHpx), and reduced glutathione (GSH) levels in renal tissues were significantly lowered in the ACR-treated groups compared to the other groups. Furthermore, histological kidney lesions indicated statistically significant differences between the groups that received ACR and the control groups. Controlling ACR-induced oxidative stress, in particular through the use of TQ, plays a crucial role in kidney protection.