Objective To explore the feasibility of establishing mini-pig model of type 1 diabetes by single intravenous injection of high dose streptozotocin. Methods Eight male Zhonghua mini-pigs (22.6±1.8 kg) were administrated with high-dose streptozotocin (150 mg/kg) into the ear vein. Before and 10, 30, 90 minutes, 1, 3 and 7 days after administration of streptozotocin, blood samples were obtained respectively, and used to dynamically monitor the fasting blood-glucose. C peptide and insulin levels were evaluated by IVGTT test. Results Since 24 hours after STZ administration, the fasting blood-glucose level was increased significantly compared with that of pre-administration and maintained at 16.7-20.6 mmol/L, while the C peptide and insulin levels were decreased significantly. IVGTT results showed that blood sugar levels at 1 h after intravenous injection of 50% glucose were much higher than 11.1 mmol/L and failed to restore to fasting glucose levels until 2 h, insulin and c-peptide did no response after injection of glucose, always kept at a trace level. Conclusion A single high-dose streptozotocin injection can be used to establish a mini-pig model of type 1 diabetes successfully.