title: Cyclopentenone isoprostanes inhibit the inflammatory response in macrophages.
abstract: Although both inflammation and oxidative stress contribute to the pathogenesis of many disease states, the interaction between the two is poorly understood. Cyclopentenone isoprostanes (IsoPs), highly reactive structural isomers of the bioactive cyclopentenone prostaglandins PGA2 and PGJ2, are formed non-enzymatically as products of oxidative stress in vivo. We have, for the first time, examined the effects of synthetic 15-A2- and 15-J2-IsoPs, two groups of endogenous cyclopentenone IsoPs, on the inflammatory response in RAW264.7 and primary murine macrophages. Cyclopentenone IsoPs potently inhibited lipopolysaccharide-stimulated IkappaB alpha degradation and subsequent NF-kappaB nuclear translocation and transcriptional activity. Expression of inducible nitric-oxide synthase and cyclooxygenase-2 were also inhibited by cyclopentenone IsoPs as was nitrite and prostaglandin production (IC50 approximately 360 and 210 nM, respectively). 15-J2-IsoPs potently activated peroxisome proliferator-activated receptor gamma (PPARgamma) nuclear receptors, whereas 15-A2-IsoP did not, although the anti-inflammatory effects of both molecules were PPARgamma-independent. Interestingly 15-A2-IsoPs induced oxidative stress in RAW cells that was blocked by the antioxidant 4-hydroxy-TEMPO (TEMPOL) or the mitochondrial uncoupler carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone. TEMPOL also abrogated the inhibitory effect of 15-A2-IsoPs on lipopolysaccharide-induced NF-kappaB activation, inducible nitricoxide synthase expression, and nitrite production, suggesting that 15-A2-IsoPs inhibit the NF-kappaB pathway at least partially via a redox-dependent mechanism. 15-J2-IsoP, but not 15-A2-IsoP, also potently induced RAW cell apoptosis again via a PPAR gamma-independent mechanism. These findings suggest that cyclopentenone IsoPs may serve as negative feedback regulators of inflammation and have important implications for defining the role of oxidative stress in the inflammatory response.
authors.affiliation: Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
authors.forename: Erik S
authors.initials: ES
authors.lastname: Musiek
authors.forename: Ling
authors.initials: L
authors.lastname: Gao
authors.forename: Ginger L
authors.initials: GL
authors.lastname: Milne
authors.forename: Wei
authors.initials: W
authors.lastname: Han
authors.forename: M Brett
authors.initials: MB
authors.lastname: Everhart
authors.forename: Dingzhi
authors.initials: D
authors.lastname: Wang
authors.forename: Michael G
authors.initials: MG
authors.lastname: Backlund
authors.forename: Raymond N
authors.initials: RN
authors.lastname: DuBois
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Zanoni
authors.forename: Giovanni
authors.initials: G
authors.lastname: Vidari
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Blackwell
authors.forename: Jason D
authors.initials: JD
authors.lastname: Morrow
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis; D016718:Arachidonic Acid / Q000737:chemistry; D001095:Arachidonic Acids / Q000737:chemistry; D015152:Blotting, Northern; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000166:cytology; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003497:Cyclic N-Oxides / Q000494:pharmacology; D051546:Cyclooxygenase 2 / Q000378:metabolism; D003517:Cyclopentanes / Q000737:chemistry*; D028441:F2-Isoprostanes / Q000737:chemistry; D017930:Genes, Reporter; D019811:Hydroxylamine; D020672:I-kappa B Proteins / Q000378:metabolism; D007249:Inflammation / Q000188:drug therapy*; D020128:Inhibitory Concentration 50; D028421:Isoprostanes / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000737:chemistry; D008264:Macrophages / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D008856:Microscopy, Fluorescence; D008956:Models, Chemical; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D009573:Nitrites / Q000737:chemistry; D018384:Oxidative Stress; D047495:PPAR gamma / Q000378:metabolism; D011453:Prostaglandins / Q000737:chemistry; D014176:Protein Biosynthesis; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D001095:Arachidonic Acids; D003497:Cyclic N-Oxides; D003517:Cyclopentanes; D028441:F2-Isoprostanes; D020672:I-kappa B Proteins; D028421:Isoprostanes; D008070:Lipopolysaccharides; D016328:NF-kappa B; C000606205:Nfkbia protein, mouse; D009573:Nitrites; D047495:PPAR gamma; D011453:Prostaglandins; D013778:Tetrazolium Salts; D013844:Thiazoles; D014409:Tumor Necrosis Factor-alpha; C108897:carbonylcyanide 4-trifluoromethoxyphenylhydrazone; D000072000:NF-KappaB Inhibitor alpha; D016718:Arachidonic Acid; D019811:Hydroxylamine; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2; C022616:thiazolyl blue; C013905:cyclopentenone; C505333:TEMPOL-H; C003959:TEMPO