Objectives: In vivo and in vitro studies have shown that 2-deoxyglucose (2-DG) administration enhances tolerance and exerts neuroprotection against ischemic injury or oxidative stress. In this study, we investigated the effects of 2-DG on ischemic brain injuries in rats and determined whether the effects are related to sublethal endoplasmic reticulum (ER) stress. Methods: 2-DG was administered systemically 7 d before the rats were subjected to focal cerebral ischemia (2 h) followed by reperfusion. Neurological score and infarct volume were evaluated, and protein expression of ER molecular chaperone glucose-regulated protein 78 (GRP78) and X-box protein-1 (XBP-1) was determined at different time points after reperfusion. Results: 2-DG pretreatment significantly decreased neurological scores after reperfusion for 3 h, 6 h, 12 h, and 24 h, reduced infarct volume at 24 h after reperfusion compared to the corresponding control groups. ER molecular chaperone GRP78 and XBP-1 increased in 2-DG pretreatment group as compared to the control. Conclusion: Pretreatment with 2-DG improves the neurological function after cerebral ischemia-reperfusion injury. Increased expression of ER chaperone GRP78 and activation of XBP-1 may contribute to the protective effect of 2-DG against brain injury.