The diagnosis of acute kidney injury (AKI) is frequently established on modifications in serum creatinine (SCr). The discriminative and prognostic aptitudes of serum cystatin-C as well as N-acetylglucosaminidase (NAG) and Interleukin-18 (IL-18) were inspected for the estimation of AKI. In this study twelve rats were alienated into two groups: control group received saline, second group received cadmium chloride at a dose (2.4 mg Cd/kg/day, i.p) for 30 days. Blood urea nitrogen (BUN), SCr, and IL-18 serum level were measured in addition to serum and tissue content of cystatin-C and NAG. AKI model showed significant increase in BUN, creatinine, and IL-18. RT-PCR showed upregulation of cystatin-C gene besides significant increase of its level in serum. Additionally, tissue content of NAG was significantly increased. Our findings may provide that grouping of several biomarkers for diagnosis of AKI is a more valuable diagnostic tool than single-marker measurement.