The role of prostaglandin E2 and nitric oxide in cell death in J774 murine macrophages.
We investigated the role of prostaglandin E2 (PGE2) and its interactions with nitric oxide (NO) on cell death and NO-mediated cytotoxicity in the murine macrophage cell line J774. Stimulation of the J774 cells with lipopolysaccharide together with interferon-gamma resulted in a dose-dependent cytotoxicity and production of PGE2 and NO, measured as nitrite. Our results showed a linear correlation between PGE2 release and cytotoxicity. The cyclooxygenase (COX) inhibitor indomethacin completely inhibited PGE2 biosynthesis, without affecting NO production or cell death. This supports previous reports suggesting that overproduction of endogenous PGE2 is mainly the consequence of cell death and does not cause it. In contrast, the NO synthase inhibitor N(omega)-monomethyl-L-arginine (L-NMMA) gave a significant, though incomplete suppression of NO release and cell death. This points to the presence of other cytotoxic factors besides NO. To evaluate the toxic effect solely due to NO, macrophages were exposed to the NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP). Incubation with SNAP also resulted in a concentration-dependent cell injury and PGE2 production. When exogenously added, PGE2 protected against SNAP-mediated cytotoxicity and simultaneously increased PGE2 release into the medium, without inducing COX-2. The cytoprotection and the stimulation of PGE2 release were both reversed by indomethacin. In conclusion, PGE2 biosynthesis may represent a mechanism by which inflammatory macrophages protect themselves against the cytotoxic effects of NO.
Division of Pharmacology (T2), Faculty of Medicine, University of Antwerp (UIA), Universiteitsplein 1, Wilryk, Belgium. zamora@uia.ua.ac.be
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D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002460:Cell Line; D015232:Dinoprostone / Q000494:pharmacology / Q000502:physiology*; D007213:Indomethacin / Q000494:pharmacology; D007371:Interferon-gamma / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D009569:Nitric Oxide / Q000494:pharmacology / Q000502:physiology*; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D019323:omega-N-Methylarginine / Q000494:pharmacology
D008070:Lipopolysaccharides; C110881:S-nitro-N-acetylpenicillamine; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D007371:Interferon-gamma; D010396:Penicillamine; D015232:Dinoprostone; D007213:Indomethacin