Objective To explore the effect of glimepiride on the glucose uptake as well as glucose transporter (GLUT)-1 and GLUT-3 expression levels of rat mandibular osteoblasts in hyperglycemia.Methods Primary osteoblasts were isolated and cultured. Then, the cells were placed in an osteogenic medium containing two glucose concentrations (5.5 and 16.5 mmol·L-1), with or without glimepiride (10 μmol·L-1). Glucose uptake was determined by employing 18F-deoxyglucose (18F-FDG) in the cells, and GLUT-1 and GLUT-3 expression levels were evaluated by Western blot analysis.Results Glucose at 16.5 mmol·L-1 significantly inhibited 18F-FDG uptake and downregulated GLUT-3 protein expression in osteoblasts. Hyperglycemia in-creased GLUT-1 protein expression. Glimepiride significantly increased glucose uptake and upregulated GLUT-1 and GLUT-3.Conclusion Glimepiride enhance the glucose transporter in rat osteoblasts at two different glucose concentrations.