title: Inhibition of LPS-stimulated NO production in mouse macrophage-like cells by azulenequinones.
abstract: Azulenequinone derivatives have been reported to display a broad spectrum of biological activities, but study at the cellular level has been limited. The effect of twenty-seven azulenequinone derivatives on nitric oxide (NO) production by mouse macrophage-like cells Raw 264.7 was investigated in this study. All of these compounds failed to stimulate the Raw 264.7 cells to produce detectable amounts of NO, but did inhibit NO production by lipopolysaccharide (LPS)-activated Raw 264. 7 cells to varying extents. Compounds [7, 8, 9, 13, 16, 25, 27], which showed lesser cytotoxic activity (CC50 = 425, 381, 482, 179, 119, 235, 225 microM, respectively), inhibited NO production to the greatest extent [selectivity index (SI) = 15.4, 26.2, 3.9, 21.6, 3.1, 6.0, 8.4, respectively]. Western blot and RT-PCR analyses demonstrated that the most active derivatives, 3-morpholino-1, 5-azulenequinone [8] and 3,7-dibromo-1, 5-azulenequinone [13], significantly reduced both the intracellular concentration of iNOS protein and the expression of iNOS mRNA. ESR spectroscopy showed that compounds [8, 13] weakly scavenged NO produced by NOC-7, possibly via their general reducing activity. These data suggest that the inhibitory effect of NO production by compounds [8, 13] might be generated mostly via the inhibition of iNOS expression, rather than the radical-mediated mechanism.
authors.affiliation: Faculty of Science, Josai University, Sakado, Saitama, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Nishishiro
authors.forename: Satomi
authors.initials: S
authors.lastname: Arikawa
authors.forename: Hidetsugu
authors.initials: H
authors.lastname: Wakabayashi
authors.forename: Ken
authors.initials: K
authors.lastname: Hashimoto
authors.forename: Kazue
authors.initials: K
authors.lastname: Satoh
authors.forename: Keiko
authors.initials: K
authors.lastname: Yokoyama
authors.forename: Senwa
authors.initials: S
authors.lastname: Unten
authors.forename: Hanzo
authors.initials: H
authors.lastname: Kakuta
authors.forename: Teruo
authors.initials: T
authors.lastname: Kurihara
authors.forename: Noboru
authors.initials: N
authors.lastname: Motohashi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sakagami
mesh_terms: D000818:Animals; D052176:Azulenes / Q000494:pharmacology*; D001565:Benzoates / Q000378:metabolism / Q000494:pharmacology; D015153:Blotting, Western; D002460:Cell Line; D003497:Cyclic N-Oxides / Q000378:metabolism / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D016166:Free Radical Scavengers / Q000494:pharmacology; D006834:Hydrazines / Q000378:metabolism / Q000494:pharmacology; D019271:Hypoxanthine / Q000378:metabolism; D007093:Imidazoles / Q000378:metabolism / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000037:antagonists & inhibitors* / Q000096:biosynthesis*; D020030:Nitric Oxide Donors / Q000378:metabolism / Q000494:pharmacology; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis / Q000235:genetics; D011809:Quinones / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013481:Superoxides / Q000378:metabolism; D014969:Xanthine Oxidase / Q000378:metabolism
chemical_list: C115239:3-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-propanamine; D052176:Azulenes; D001565:Benzoates; D003497:Cyclic N-Oxides; D016166:Free Radical Scavengers; D006834:Hydrazines; D007093:Imidazoles; D008070:Lipopolysaccharides; D020030:Nitric Oxide Donors; D011809:Quinones; D012333:RNA, Messenger; D013481:Superoxides; C079393:1,3-dihydroxy-4,4,5,5-tetramethyl-2-(4-carboxyphenyl)tetrahydroimidazole; D019271:Hypoxanthine; D009569:Nitric Oxide; C017245:5,5-dimethyl-1-pyrroline-1-oxide; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D014969:Xanthine Oxidase