Orazipone inhibits activation of inflammatory transcription factors nuclear factor-kappa B and signal transducer and activator of transcription 1 and decreases inducible nitric-oxide synthase expression and nitric oxide production in response to inflammatory stimuli.
Orazipone [OR-1384; 3-[4-(methylsulfonyl)benzylidene]pentane-2,4-dione] is a novel sulfhydryl-modulating compound that has anti-inflammatory properties in experimental models of asthma and inflammatory bowel disease. In inflammation, inducible nitricoxide synthase (iNOS) generates NO, which modulates the immune response. Compounds that inhibit iNOS expression or iNOS activity possess anti-inflammatory effects. In the present study, we examined the effects of orazipone and its derivative OR-1958 [3-[3-chlorine-4-(methylsulfonyl)benzylidene]pentane-2,4-dione] on iNOS expression and NO production in J774 macrophages stimulated by bacterial lipopolysaccharide (LPS) and in human alveolar epithelial cells activated by proinflammatory cytokines. Protein expression and nuclear translocation of transcription factors were measured by Western blot. iNOS mRNA expression was determined by quantitative reverse transcription-polymerase chain reaction and iNOS mRNA stability by actinomycin D assay. iNOS promoter activity was studied in a cell line expressing luciferase under the control of iNOS promoter. Orazipone and its derivative OR-1958 but not its nonthiol-modulating analog inhibited iNOS expression and NO production in a concentration-dependent manner. Orazipone decreased LPS-induced iNOS mRNA expression, but the decay of iNOS mRNA was not affected. Orazipone extensively prevented LPS-induced activation of nuclear factor kappaB (NF-kappaB) and signal transducer and activator of transcription (STAT) 1, which are important transcription factors for iNOS. In agreement, human iNOS promoter activity was inhibited by orazipone. In conclusion, orazipone decreased activation of inflammatory transcription factors NF-kappaB and STAT1, and expression of iNOS in cells exposed to inflammatory stimuli. The thiolmodulating property seems to be critical in mediating the antiinflammatory effects of orazipone.
The Immunopharmacology Research Group, Medical School, University of Tampere, Tampere FIN-33014, Finland.
Outi
O
Sareila
Mari
M
Hämäläinen
Erkki
E
Nissinen
Hannu
H
Kankaanranta
Eeva
E
Moilanen
D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology; D006801:Humans; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D009569:Nitric Oxide / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D050794:STAT1 Transcription Factor / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000737:chemistry / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology*
D016328:NF-kappa B; D050794:STAT1 Transcription Factor; D013438:Sulfhydryl Compounds; D009569:Nitric Oxide; C502617:orazipone; D052247:Nitric Oxide Synthase Type II