The goal of the present study was to evaluate the antioxidant and anti-apoptotic roles of sesame oil (SO) and N acetylcysteine (NAC) against gentamicin (GM) induced nephrotoxicity. This study was carried out on 36 male Wistar rats divided into six equal groups: control (saline, once daily, PO); SO group (5 mlof SO/kg, once daily, PO); NAC group (150 mg of NAC/kg, once daily, PO); GM group (80 mgof GM/kg, IP). The SO+GM and NAC+GM groups received SO, NAC as mentioned. Saline, SO, and NAC were administered for 30 days, while GM was given one hour after SO and NAC administration in the last 9 days of the experiment. Levels of urea, creatinine, and uric acid were significantly increased in GM-treated groups when compared to control. The concentration of malondialdehyde (MDA) in the kidney tissues of the GM-treated groups were notably substantially higher than in the control group. Superoxide dismutase (SOD) and reduced glutathione (GSH) levels in renal tissues were significantly lower in the GM-treated groups compared to the control group. Furthermore, the microscopic kidney lesions showed adifference between the groups that received GM and the control groups. Additionally, GM increased the expression of Tumor Necrosis Factor α (TNF-α), Bax, and decreased the expression of Bcl2. When compared with the effect ofSO, NAC had a better effect on controlling GM damage in the kidneys. Controlling GM-induced oxidative stress, in particular through the use of SO and NAC, plays a crucial role in kidney protection.