The hypoglycemic effect of mulberry leaves using alloxan-induced diabetic mice was investigated. The hypoglycemic effect of mulberry leaves was significantly affected with anaerobic incubation up to for 6 hrs. The body weight of mice treated with 20% anaerobic conditioned mulberry leaves was higher than that in the control group. Mulberry leaves which were treated with anaerobic condition significantly decreased blood glucose level in alloxan-induced type I diabetes mice. Furthermore, glucose tolerance was significantly recovered by feeding mulberry leaves anaerobically treated. Pathological analysis revealed that the disappearance of insulin-secreting beta-cells in the islets of Langerhans from the alloxan induced diabetic mice was strongly inhibited in the mulberry leaves feeding group. The content of 1-deoxynojirimycin increased up to 5% in detached mulberry leaves incubated under nitrogen gas for 6 hrs. Nevertheless, it is not clear whether the synthesis of 1-deoxynojirimycin is increased or the degradation of it is decreased. Overall, results obtained from this study indicate that the hypoglycemic effect was increased by the mulberry leaves with anaerobic treatment.