Previous studies utilizing an ex vivo porcine model of intestinal ischemic injury demonstrated that PGE₂ stimulates repair of mucosal barrier function via a mechanism involving chloride (Cl^-) secretion and reductions in paracellular permeability. Further experiments revealed that the signaling mechanism for PGE₂-induced mucosal recovery was mediated via type-2 chloride channels (ClC-2). Therefore, the objective of the present study was to directly investigate the role of ClC-2 in mucosal repair by evaluating mucosal recovery in ischemic-injured intestinal mucosa treated with the selective ClC-2 agonist, lubiprostone. Ischemic-injured porcine ileal mucosa was mounted in Ussing chambers and short circuit current (I_sc) and transepithelial electrical resistance (TER) were measured in response to lubiprostone. Application of 0.01µM - 1µM lubiprostone to ischemic-injured mucosa induced concentration-dependent increases in TER, with 1µM lubiprostone stimulating a 2-fold increase in TER (TER = 26 ·cm², p<0.01). Furthermore, lubiprostone significantly (p<0.05) reduced mucosal-to-serosal fluxes of ³H-mannitol to levels comparable to those of normal control tissues, and restored occludin localization to tight junctions. Activation of ClC-2 with the selective agonist, lubiprostone, stimulated elevations in TER and reductions in mannitol flux in ischemic-injured intestine associated with structural changes in tight junctions. Prostones such as lubiprostone may provide a selective and novel pharmacological mechanism of accelerating recovery of acutely injured intestine as compared to the non-selective action of prostaglandins such as PGE₂.