<jats:title>Abstract</jats:title><jats:p>Acute ethanol intoxication and exposure (AE) has been known to impair wound healing and associated angiogenesis. Here, we found that AE diminished the formation of novel reparative lipid mediator 14<jats:italic>S</jats:italic>,21‐dihydroxy‐docosa‐4<jats:italic>Z</jats:italic>,7<jats:italic>Z</jats:italic>,10<jats:italic>Z</jats:italic>,12<jats:italic>E</jats:italic>,16<jats:italic>Z</jats:italic>,19<jats:italic>Z</jats:italic>‐hexaenoic acid (14<jats:italic>S</jats:italic>,21‐diHDHA) and its biosynthetic intermediate 14<jats:italic>S</jats:italic>‐hydroxy‐DHA (14<jats:italic>S</jats:italic>‐HDHA) from docosahexaenoic acid (DHA) in murine wounds. However, AE did not reduce the formation of DHA and the intermediate 21‐HDHA. These results indicate that in the biosynthetic pathways of 14<jats:italic>S</jats:italic>,21‐diHDHA in wounds, AE suppresses the 14<jats:italic>S</jats:italic>‐hydroxy‐generating activity of 12‐lipoxygenase‐like (LOX‐like), but does not suppress the 21‐hydroxy‐generating activity of cytochrome P450 and DHA‐generating activities. The AE‐suppression of 12‐LOX‐like activity was further confirmed by the diminished formation of 12‐hydroxy‐eicosatetraenoic acid in wounds under AE. Supplementing 14<jats:italic>S</jats:italic>,21‐diHDHA to wounds rescued the AE‐impaired healing and vascularization. 14<jats:italic>S</jats:italic>,21‐diHDHA restored AE‐impaired processes of angiogenesis <jats:italic>in vitro</jats:italic>: endothelial cell migration, tubulogenesis, and phosphorylation of p38 mitogen‐activated protein kinase (MAPK). Taken together, the suppression of 14<jats:italic>S</jats:italic>,21‐diHDHA formation is responsible, at least partially, for the AE‐impairment of cutaneous wound healing and angiogenesis. Supplementing 14<jats:italic>S</jats:italic>,21‐diHDHA to compensate its deficit in AE‐impaired wounds rescues the healing and angiogenesis. These results provide a novel mechanistic insight for AE‐impaired wound healing that involves the necessary roles of 14<jats:italic>S</jats:italic>,21‐diHDHA. They also offer leads for developing 14<jats:italic>S</jats:italic>,21‐diHDHA‐related therapeutics to ameliorate AE‐impairment of wound healing. J. Cell. Biochem. 111: 266–273, 2010. © 2010 Wiley‐Liss, Inc.