Abstract
Lipoxin A(4) (LXA(4)) and aspirin-triggered 15-epi-LXA(4) (ATL) are emerging as endogenous braking signals for neutrophil-mediated tissue injury. Recent studies indicate that peroxynitrite (ONOO(-)) may function as an intracellular signal for the production of IL-8, a potent proinflammatory cytokine in human leukocytes. In this study, we evaluated the impact of the metabolically stable analogues of LXA(4)/ATL on lipopolysaccharide (LPS)-induced ONOO(-) formation and ONOO(-)-mediated IL-8 gene expression in human leukocytes. At nanomolar concentrations, LXA(4) analogues markedly reduced LPS-stimulated superoxide formation, evoked increases in intracellular diamino-fluorescein fluorescence (an indicator of NO formation), and consequently reduced ONOO(-) formation in isolated neutrophils, as well as in neutrophils, monocytes, and lymphocytes, in whole blood. LXA(4)/ATL analogues attenuated nuclear accumulation of activator protein-1 and nuclear factor-kappaB in both polymorphonuclear and mononuclear leukocytes and inhibited IL-8 mRNA expression and IL-8 release by 50-65% in response to LPS. The LXA(4) inhibitory responses were concentration dependent and were not shared by 15-deoxy-LXA(4). None of the LXA(4) analogues studied affected neutrophil survival, nor reversed the apoptosis delaying action of LPS in neutrophils. In addition, LXA(4) analogues had no significant effect on exogenous ONOO(-)-induced IL-8 gene and protein expression. These findings suggest that by attenuating ONOO(-) formation, LXA(4) and ATL can oppose ONOO(-) signaling in leukocytes and provide a rationale for using stable synthetic analogues as antiinflammatory compounds in vivo.