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Inflamm Res ; 50(8): 409-14, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11556521

RESUMO

OBJECTIVE AND DESIGN: To evaluate the capacity of doxycycline and minocycline to inhibit NO production and N-nitrosation reactions in vitro. METHODS: Synovial cells obtained from 6 patients with osteoarthritic joint disease were incubated for 24 hours with (i) or without (ii) IL-1beta (1 ng/ml), TNF-alpha (500 pg/ml), IFN-gamma (10(4) U/ml) plus minocycline or doxycycline (10(-4) to 10(-6) M), diclofenac (10(-5) M), or cortisol (10(-5) M). Nitrosothiols were determined by fluorimetry, nitrite by the Griess reaction, nitrate by a spectrophotometric assay using oxidation by nitrate reductase and iNOS by immunoblotting. RESULTS: After 24 hours of stimulation, the level of NO production was much higher than that in untreated cells: about 5.5 times higher for nitrosothiols, 5.2 times higher for nitrate and about 3.5 times higher for nitrite. Doxycycline and minocycline induced a dose-dependent decrease in the production of nitrosothiols, nitrate and nitrite, and inhibited the synthesis of the iNOS protein. Doxycycline and minocycline inhibited the N-nitrosation reaction of DAN effectively, with IC50 values close to 100 microM. Diclofenac and cortisol had no effect. CONCLUSION: This study provides new information on the mechanism by which tetracyclines exert anti-inflammatory effects, via inhibiting nitrosothiols.


Assuntos
Citocinas/farmacologia , Osteoartrite/patologia , S-Nitrosotióis/metabolismo , Membrana Sinovial/metabolismo , Tetraciclinas/farmacologia , Antibacterianos/farmacologia , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios não Esteroides/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Doxiciclina/farmacologia , Humanos , Immunoblotting , Minociclina/farmacologia , Nitratos/metabolismo , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo II , Nitritos/metabolismo , Ovalbumina/farmacologia , Inibidores de Serina Proteinase/farmacologia , Esteroides , Membrana Sinovial/citologia
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