Kidney function is maintaining of homeostasis and toxins elimination. Thus, it is susceptible to acute damage by xenobiotics. Drug-induced nephrotoxicity is becoming a principal etiology for acute kidney injury (AKI). The present study was designed to investigate the protective effect of iron chelator, desferroxamin (DFO), and hesperidin (HES) on gentamicin (GM) induced AKI in rats and elucidating the potential mechanisms. Group I: control, group II: GM (100mg/kg/d. i.p.) for 7days, group III: HES (200 mg/ kg) two days before and 7 days concomitantly with GM, group IV: DFO (100 mg/kg/d. i.p.) coadministrated with GM and group V: GM + combined treatment by DFO and HES. Serum urea, creatinine, KIM1, TNF-α and IL-6, renal MDA and HO1 concentration, immunohistochemical expression of caspase 3 and histopathology of kidney were evaluated. GM induced significant increase in all parameters excluding renal HO1 level which is significantly decreased (p