Diabetes mellitus (DM) is a major cause of morbidity and mortality worldwide, and its complications are prominent public health issues. Many experimental models of streptozotocin (STZ)-induced and high-fat diet (HF)-induced DM have been used to study this disease. Studies have indicated that unilateral nephrectomy (UN) accelerates the development of diabetic nephropathy. We hypothesized that UN stimulates HF and STZ combination-induced DM in mice. Seventy-two female C57BL/6J mice were divided into four treatment groups: HF; HF + STZ120 (HF and STZ, 120 mg/kg); UN + HF + STZ120 (UN, HF and STZ, 120 mg/kg); and HF + STZ200 (HF and STZ, 200 mg/kg). Onset of DM, survival rate, blood pressure, urine glucose level, and pancreatic histology were investigated. Additionally, renal function was evaluated in the UN + HF + STZ120 group after STZ injection. DM was induced in the UN + HF + STZ120 and HF + STZ200 groups within one week. The UN + HF + STZ120 group had lower mortality than the HF + STZ200 group and greater pancreatic destruction than the HF and HF + STZ120 groups. Two weeks after STZ injection, blood pressure was not significantly different among the groups. Nephrotoxicity associated with the combination of UN and STZ was not observed. In conclusion, the combination of these three techniques - UN, HF and STZ induced DM rapidly and effectively.