Objective The aim of this study was to observe pregestational ethinyl estradiol (EE)-exposure-induced glucose metabolism alterations and hepatic glucose-metabolism-related gene expression changes in the offsprings of SD rats. Methods Fifty-two female and 52 male SD rats were used in this study. The female rats were gavaged with sesame oil, 50 μg/kg, 200 μg/kg and 800 μg/kg EE for 15 consecutive days. After the end of exposure period, the female rats were mated with male rats and gave birth to next generation. The blood glucose and insulin tolerance in the offsprings were measured on postnatal days 23 (P23) and 25 (P25). The expressions of hepatic glucose-metabolism-related genes were measured by RT-PCR. Results In the female offsprings, the 200 μg/kg EE group had significantly lower fasting blood glucose level than that in the control group (P < 0.05). At 15 min after glucose administration, the blood glucose level in the 800 μg/kg EE group was much higher than that in the control group, 50 μg/kg and 200 μg/kg EE groups (P < 0.01, P < 0.01, P < 0.01). At the time point of 2 h, the blood glucose level of the 50 μg/kg and 800 μg/kg EE groups were both significantly lower than that in the control group (P < 0.05, P < 0.01). The female offsprings in the 50 μg/kg and 200 μg/kg EE groups had significantly higher glucose level after insulin administration than that in the control group (P < 0.001, P < 0.01). In the male offsprings, the 800 μg/kg EE group had a significantly higher blood glucose level than the control group at 15 min after glucose administration (P < 0.01), and the 200 μg/kg EE group had a lower blood glucose level than the control at 30 min after glucose administration. In the male offsprings, the blood glucose level of 50 μg/kg and 200 μg/kg EE groups were much higher than that of the control group (P < 0.01, P < 0.01). In the female offsprings, Glut2 and Lpk mRNA expressions in the 50 μg/kg, 200 μg/kg and 800 μg/kg EE groups were much lower than that in the control group (P < 0.01, P < 0.05, P < 0.05). Gys2 mRNA expressions in the 50 μg/kg and 200 μg/kg EE groups were much lower than that in the control group (P < 0.01, P < 0.01). In the male offsprings, the 200 μg/kg EE group had much higher G6pase and Pepck mRNA expression than in the control group (P < 0.01, P < 0.01).The Glut2 mRNA expression in the 50 μg/kg EE group was much lower than that in the control group (P < 0.01). The Gys2 mRNA expression in the 800 μg/kg EE group was significantly higher than that in the control group (P < 0.01). Conclusions Pregestational EE exposure can lead to impaired glucose tolerance and insulin resistance in female offspring and alterations of key hepatic glucose-metabolism-related gene expression, and these effects are sex-specific,and female offspring is more sensitive to pregestational EE exposure.