Inhibition of LPS-stimulated NO production in mouse macrophage-like cells by azulenes.
We investigated the effect of twenty-seven azulenes on nitric oxide (NO) production by mouse macrophage-like Raw 264.7 cells. No azulene derivative alone induced NO production by the Raw 264.7 cells, but inhibited lipopolysaccharide (LPS)-stimulated NO production to various extents. The ability of azulenes to inhibit NO generation by activated macrophages was generally increased when their cytotoxic activity declined. Western blot and RT-PCR analyses demonstrated that the most potent compound, 1,3-difluoroazulene [11], slightly inhibited the expression of inducible NO synthase (iNOS), but only at extremely high concentrations. ESR spectroscopy showed that [11] did not produce radical under alkaline condition, nor scavenged O2- (generated by HX-XOD reaction) or NO (generated by NOC-7). These data suggest that the inhibitory effect of [11] may be produced via a mechanism other than iNOS induction and a radical-mediated mechanism.
Hashiba
Kana
K
Faculty of Science, Josai University, Sakado, Saitama, Japan.
Yokoyama
Keiko
K
Wakabayashi
Hidetsugu
H
Hashimoto
Ken
K
Satoh
Kazue
K
Kurihara
Teruo
T
Motohashi
Noboru
N
Sakagami
Hiroshi
H
D000818:Animals; D052176:Azulenes; D001565:Benzoates / Q000737:chemistry; D015153:Blotting, Western; D002460:Cell Line; D003508:Cycloheptanes / Q000737:chemistry / Q000494:pharmacology*; D004578:Electron Spin Resonance Spectroscopy; D016166:Free Radical Scavengers / Q000494:pharmacology; D006834:Hydrazines / Q000737:chemistry; D007093:Imidazoles / Q000737:chemistry; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D008262:Macrophage Activation / Q000187:drug effects / Q000502:physiology; D008264:Macrophages / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D051379:Mice; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019097:Spin Trapping; D013329:Structure-Activity Relationship
C115239:3-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-propanamine; D052176:Azulenes; D001565:Benzoates; D003508:Cycloheptanes; D016166:Free Radical Scavengers; D006834:Hydrazines; D007093:Imidazoles; D008070:Lipopolysaccharides; D012333:RNA, Messenger; C079393:1,3-dihydroxy-4,4,5,5-tetramethyl-2-(4-carboxyphenyl)tetrahydroimidazole; D009569:Nitric Oxide; C005525:azulene; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse