Objective To explore a gestational insulin resistance mice model establishing method, which is steady, reliable and similar to human being. Method Female KM mice at 5 weeks were randomly divided into high fat diet group(n=30) and normal diet group(n=30). High fat diet group was exposed to high diet for 4 weeks, and then male and female were mated(n:n=1:1) Then the pregnancy mice were intraperitoneal injected with 30 mg/kg/d STZ during the first 3 days, the control group was injected with equal dosage of citrate buffer solution(0.1 mol/L, pH=4.2) for 3 days. Their blood glucose, body weight, the amount of food intake and water intake were recorded at 3, 7, 14 and 19 d of the GDM model respectively. The serum INS, ADP, LEP, CRP were measured by ELISA. Results We successfully made the GDM mice model, and the pregnant mice showed significant signs of polyuria, polydipsia, hyperphagia and weight loss compared to the control model(P < 0.01). The blood glucose of GDM mice >11.1mmol/L, the serum cytokines (INS(1.50±0.25)Mu/L, ADP(0.65±0.13)μg/L, LEP(1.60±0.12)μg/L, CRP(37.54±4.70)μg/L) of GDM mice were significant difference compared to the control model(P < 0.01). Postpartum, the GDM mice blood glucose returned to the normal level. Conclusion Gestational diabetes mellitus mice model can be successfully developed by high fat diet with low dose STZ and three times induce, which preferable mimics the characteristic of gestational pathologic insulin resistance in human beings.