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 ml of SO/kg, once daily, PO); NAC group (150 mg of NAC/kg, once daily, PO); GM group (80 mg of 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 the control. The concentration of malondialdehyde (MDA) in the kidney tissues of the GM-treated groups was 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 a difference between the groups that received GM and the control groups. Additionally, GM increased the expression of Tumor Necrosis Factor α (TNFα), Bax, and decreased the expression of Bcl2. When compared with the effect of SO, NAC had a better effect on controlling GM damage in the kidneys. Controlling GM-induced oxidative stress, in particular using SO and NAC, plays a crucial role in kidney protection.