Fipronil (FPN) is phenylpyrazole insecticide extensively used to control a wide variety of pests. Betanin (BET) is a natural colorant with promising antioxidant and anti-inflammatory effects. This study aimed to investigate the potential protective effect of BET on FPN induced nephrotoxicity in adult male albino rats. Forty rats were assigned into 4 equal groups; Group I (Control); Group II (BET) received 20 mg/kg b.wt/day; Group III (FPN) received 4.8 mg/kg b.wt/day; and Group IV (BET/FPN). All treatments were given orally for 90 days. At the end of experiment, blood samples were collected for analysis of serum urea and creatinine. Kidneys were harvested for determination of kidney injury molecule-1(KIM-1) level; gene expression of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and NAD(P)H: quinone oxidoreductase-1 (NQO-1); oxidative stress biomarkers including malondialdehyde (MDA), protein carbonyl content (PCC), catalase activity (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH). Histopathological examination and immunohistochemical investigation of Nrf2, nuclear factor kappa B (NF-κB), and caspase-3 were also undertaken. The results revealed kidney dysfunction, downregulation of Nrf2, HO-1, and NQO-1 genes, redox imbalance, structural damage, decreased Nrf2 and increased NF-κB immune-expression, in addition to strong caspase-3 immunoreactivity in FPN-treated group. In the combined group, BET co-administration resulted in functional and structural amelioration, up-regulation of Nrf2, HO-1, and NQO-1 genes, mitigation of redox imbalance, and strong anti-inflammatory and antiapoptotic effects. In conclusion, BET via activation of Nrf2-HO-1/NQO-1 pathway, exhibits beneficial antioxidant, anti-inflammatory, and antiapoptotic effects against FPN-induced nephrotoxicity.