The receptor for advanced glycation end-products (RAGE) has been implicated in the pathogenesis of numerous conditions associated with excessive inflammation. To determine whether RAGE-dependent signaling is important in the development of intestinal barrier dysfunction after hemorrhagic shock and resuscitation (HS/R), C57Bl/6, rage^-/- or congenic rage ^+/+ mice were subjected to HS/R (mean arterial pressure of 25 mm Hg for 3 h) or a sham procedure. Twenty-four h later, bacterial translocation (BT) to mesenteric lymph nodes and ileal mucosal permeability to FITC-labeled dextran were assessed. Additionally, samples of ileum were obtained for immunofluorescence microscopy and plasma was collected for measuring IL-6 and IL-10 levels. HS/R in C57Bl/6 mice was associated with increased BT, ileal mucosal hyperpermeability and high circulating levels of IL-6. All of these effects were prevented when C57Bl/6 mice were treated with recombinant human sRAGE (the extracellular ligand-binding domain of RAGE). HS/R induced BT, ileal mucosal hyperpermeability and high plasma IL-6 levels in rage ^+/+ but not rage^-/- mice. Circulating IL-10 levels were higher in rage^-/- as compared to rage ^-/- mice. These results suggest that activation of RAGE-dependent signaling is a key factor leading to gut mucosal barrier dysfunction after HS/R.