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Background of the Study: The increase in the therapeutic use of tramadol in the management of moderate to severe pains in some disease conditions and its unregulated access has led to its associated toxicity and there is little or no information on the protection against its associated toxicity., Aim of the Study: Considering the medicinal value of pumpkin seed oil, its availability, and neglected use, it becomes necessary to evaluate the possible potential of the seed oil in tramadol-induced oxidative stress in Wister Albino rats., Methods of the Study: This study used fifty-six (56) albino rats to determine the impact of Cucurbita pepo seed oil (CPSO) on tramadol-induced oxidative stress. The rats were grouped into 7. After a week of acclimatization, rats in group 1 (normal control) had access to water and food, while rats in group 2 received 5 mL/Kg (b.w) of normal saline. 100 mg/kg of tramadol (TM) was delivered to groups 3-6 to induce toxicity. The third group (TM control) received no treatment, whilst the other 3 groups (TM-CPSO treatment groups) received 5, 2.5, and 1.5 mL/Kg of CPSO, respectively. Group 7 received only 5 mL/kg CPSO (CPSO group). Similarly, groups 2 through 7 had unrestricted access to food and water for 42 days and received treatments via oral intubation once per day. Indicators of oxidative stress were discovered in the brain homogenate., Results: TM toxicity was demonstrated by a considerable increase ( P < .05) in the brain MDA level and a significant drop ( P < .05) in the brain GSH level, as well as a significant reduction ( P < .05) in GPx, catalase, SOD, GST, and quinone reductase activities., Conclusion: The dose-dependent delivery of CPSO was able to restore not only the activity but also the concentrations of the altered markers., Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2024.)