BLT2, a low-affinity leukotriene B4 (LTB4) receptor, is highly expressed in intestinal epithelial cells. Recently, 12(S)-hydroxyheptadeca-5Z,8E,10E-trienoic acid (12-HHT) was identified as an endogenous ligand for BLT2. However, the precise role of this receptor has not been fully understood. The present study investigated the role of BLT2 in the healing of intestinal injury using BLT2-deficient (BLT2KO) mice, transgenic mice with intestinal epithelium-specific overexpression of BLT2 (villin-BLT2-Tg), and murine epithelial cell line (YAMC). The intestinal injury was induced in mice by subcutaneous administration of indomethacin and the healing of injury was determined 48, 72 and 96 h later. The wound (diameter: 1 mm) was inflicted on monolayer of YAMC and wound closure was evaluated 6 h later. Further, cell proliferation was determined by WST-1 assay. The healing of indomethacin-induced intestinal injury was significantly delayed in BLT2KO mice when compared with wild-type (WT) mice. In contrast, villin-BLT2-Tg mice exhibited healing-promoting properties when compared with WT mice. In YAMC, CAY10583, a BLT2 agonist, concentration-dependently promoted wound repair and cell proliferation. The similar effect was observed by 12-HHT. These findings suggest that BLT2 accelerates the healing of intestinal injury. This effect is at least partly mediated via promotion of epithelial cell proliferation.