Objective: To investigate the effect of glucagon-like peptide 2 (GLP-2) on bacterial translocation and endotoxemia in rats with bile duct ligation. Methods: Seventy-two SD rats were equally randomized into sham operation group, model group and GLP-2 treatment group, and rats in model and GLP-2 treatment group underwent common bile duct ligation. After surgery, rats of the GLP-2 treatment group were subjected to daily intraperitoneal injection of GLP-2 [250 μg/ (kg?d)], and those of the sham operation group and model group received the same volume of PBS by using exactly the same regimen. The phosphatidylinositol 3-kinase (PI3K) expression in the intestinal mucosa, and the bacterial translocation rate and development of endotoxemia of the rats were detected at day 1, 3, and 7 after surgery. Results: The PI3K expressions in the intestinal mucosa of rats of the model group decreased significantly at day 3 and 7 after surgery compared with those of the sham operation group or GLP-2 treatment group (all P<0.05), and although those somewhat decreased in the GLP-2 treatment group, the differences had no statistical significance compared with sham operation group (all P>0.05). In the model group, the rates of bacterial translocation to the liver and spleen increased gradually and both reached 100% on day 7 after surgery. The overall rates of bacterial translocation to the two organs were 75.0% and 66.7%, respectively. In the GLP-2 treatment group, the rates of bacterial translocation to the two organs increased slowly, and the overall rates of bacterial translocation were 37.5% and 25.0% respectively, which were significantly lower than those in the GLP-2 treatment group (both P<0.05). The endotoxin content of the portal vein blood elevated gradually with time in both model group and GLP-2 treatment group, but it was significantly lower in GLP-2 treatment group than that in model group at each observation point (all P<0.05). Conclusion: GLP-2 has inhibitory effect on the bacterial translocation from the intestinal tract and development of endotoximia in rats with common bile duct ligation, and the mechanism is probably related to its enhancement of PI3K expression in intestinal mucosa and thereby protecting gut barrier function.