Aflatoxin B (AFB) is a toxic food pollutant that has extensive deleterious impacts on the kidney. Oxidative stress represents the primary mechanism of AFB nephrotoxicity and can also cause mitochondrial damage. Damaged mitochondria can trigger apoptosis leading to kidney injury. PINK1/Parkin-mediated mitophagy can alleviate mitochondrial injury to maintain cellular homeostasis, however, its role in AFB-induced kidney damage is unknown. To investigate the effect of PINK1/Parkin-mediated mitophagy on kidney impairment triggered by AFB, 40 male wild-type (WT) C57BL/6N mice were first assigned to 4 groups and orally exposed to AFB at 0, 0.5, 0.75, and 1 mg/kg body weight (BW) for 28 days. The results revealed that AFB induced kidney damage, oxidative stress, mitochondrial damage, apoptosis and activated PINK1/Parkin-mediated mitophagy with a dose-dependent effect. Then, 20 male WT C57BL/6N mice and 20 male Parkin knockout (Parkin) C57BL/6N mice were assigned to 4 groups and orally exposed to AFB at 0, 1, 0, and 1 mg/kg BW for 28 days. The results revealed that Parkin suppressed mitophagy and exacerbated kidney damage, oxidative stress, mitochondrial damage, and apoptosis under AFB exposure. The aforementioned evidences demonstrate that PINK1/Parkin-mediated mitophagy is activated by AFB and protects against kidney damage in mice.