Empagliflozin (EMPA) is a glucose-lowering agent that is widely used for treatment of diabetes mellitus and diabetic nephropathy. This work aimed to evaluate the therapeutic and prophylactic effects of EMPA and NAC in treating nephrotoxicity caused by 5-Fluorouracil (5-FU) in male Albino Wistar rats. 48 rats were divided into 6 groups: The first group used as negative control. The second received 20 mg/kg of 5-FU intraperitoneally for 6 days. The third group received 5-FU plus 10 mg/kg EMPA orally for 10 days. The fourth group received EMPA 10 days before the injection of 5-FU. The fifth group received 5-FU plus N-acetylcysteine (NAC) 40 mg/kg. The sixth group received 5-FU, NAC plus EMPA. Biochemical evaluation for urea, creatinine, uric acid, albumin, TNF-α and IL-1β in serum and GSH and MDA in renal homogenate were done. Histopathological examination of kidneys was done with immunohistochemical analysis of caspase-3. The nephrotoxic effect of 5-FU was characterized by elevation of creatinine, urea, uric acid, MDA, TNF-α and IL-1β with reduction of albumin and GSH. EMPA caused improvement in kidney status especially when used therapeutically or with NAC. The immunohistochemical analysis showed that EMPA caused a reduction in the expression of caspase-3. EMPA is an effective drug in cases of 5-FU induced nephrotoxicity. It is more effective when used as a treatment rather than a prophylactic strategy. The effect of EMPA is enhanced when combined with NAC. EMPA nephroprotective effect is mediated via antioxidant, antiinflammatory effects and by decreasing the expression of renal caspase-3.