The omega-3 fatty acid docosahexaenoate attenuates endothelial cyclooxygenase-2 induction through both NADP(H) oxidase and PKC epsilon inhibition

A high intake of the omega-3 fatty acid docosahexaenoate [doco- sahexaenoic acid (DHA)] has been associated with systemic anti- inflammatory effects and cardiovascular protection. Cyclooxygen- ase (COX)-2 is responsible for the overproduction of prostaglandins (PG) at inflammatory sites, and its expression is increased in atheroma. We studied the effects of DHA on COX-2 expression and activity in human saphenous vein endothelial cells challenged with proinflammatory stimuli. A >24-h exposure to DHA reduced COX-2 expression and activity induced by IL-1, without affecting COX-1 expression. DHA effect depended on the NF- B-binding site in the COX-2 promoter. EMSAs confirmed that DHA attenuated NF- B activation. Because MAPK, PKC, and NAD(P)H oxidase all partici- pate in IL-1-mediated COX-2 expression, we also tested whether these enzymes were involved in DHA effects. Western blots showed that DHA blocked nuclear p65 NF- B subunit translocation by decreasing cytokine-stimulated reactive oxygen species and ERK12 activation by effects on both NAD(P)H oxidase and PKC activities. Finally, to address the question whether DHA itself or DHA-derived products were responsible for these effects, we inhibited the most important enzymes involved in polyunsaturated fatty acid metabolism, showing that 15-lipoxygenase-1 products mediate part of DHA effects. These studies provide a mechanistic basis for antiinflammatory and possibly plaque-stabilizing effects of DHA