Objective To explore the effect of ghrelin on insulin secretion and expression of glucose transporter protein-2 (Glut-2) in isolated pancreas of rats. Methods Twenty five Wistar rats were randomly devided into normal control group (NC group), high concentration of glucose group (HCG group), high concentration of glucose with high concentration of ghrelin group (10-8mol/L, HCG+HCGh group), medium concentration of ghrelin group(10-9mol/L, HCG+MCGh group), and low concentration of ghrelin group (10-10mol/L, HCG+LCGh group) with 5 rats in each group. The rat isolated pancreas perfusion models were established firstly, then from the distal end of abdominal aortas, the models were perfused with low concentration of glucose (5.5mmol/L), high concentration of glucose (33.3mmol/L) or high concentration of glucose added with different concentrations of ghrelin. Levels of insulin outflowed from portal vein were tested by ELISA method, expression levels of Glut-2 protein were tested by immunohistochemical method,and ultrastructure changes of islet β cell were observed under the transmission electron microscope. Results There were no significant difference on levels of fasting blood glucose (FBG), fasting insulins (FINS), homeostasis model of assess-ment for insulin resistence index (HOMA-IR), and homeostasis model of assessment for pancreatic β cell function (HOMA-β),(P>0.05). There were no significant difference on insulin levels of effluent from portal vein of 5 groups (P>0.05) when isolated pancreas perfused with 5.5mmol/L glucose, while had 2 secretion peaks in 3min and 10-12min after 33.3 mmol/L glucose perfusion, where HCG+HCGh group at the top. The mean density value of Glut-2 protein in NC group was higher than that of other 4 groups (P<0.05). The results of transmission electron microscopy showed that apoptosis was lighter in NC group than that of other 4 groups, and apoptosis of HCG+HCGh group was lighter than that of HCG+MCGh group and HCG+LCGh group. Conclusions In isolated pancreas of rats, ghrelin promotes high concentration of glucose-stimulated insulin secretion, decreases expression of Glut-2 protein, and protects the islet β cell.