Synergistic induction of nitric oxide synthase type II: in vitro effect of leptin and interferon-gamma in human chondrocytes and ATDC5 chondrogenic cells.

Imagen de Miguel Otero
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TítuloSynergistic induction of nitric oxide synthase type II: in vitro effect of leptin and interferon-gamma in human chondrocytes and ATDC5 chondrogenic cells.
Publication TypeJournal Article
Year of Publication2003
AutoresOtero, M, Reino, JJesús Gom, Gualillo, O
JournalArthritis Rheum
Volume48
Issue2
Pagination404-9
Date Published2003 Feb
ISSN0004-3591
Palabras claveAnimals, Antineoplastic Agents, Cartilage, Articular, Cell Line, Chondrocytes, Drug Synergism, Gene Expression Regulation, Enzymologic, Humans, In Vitro Techniques, Interferon-gamma, Janus Kinase 2, Leptin, Mice, Nitric Oxide, Nitric Oxide Synthase, Nitric Oxide Synthase Type II, Protein-Tyrosine Kinases, Proto-Oncogene Proteins
Abstract

OBJECTIVE: To study, in vitro, the effect of leptin (OB), alone or in combination with interferon-gamma (IFNgamma), on inducible nitric oxide synthase (iNOS) and NO production in human primary chondrocytes and in mouse embryonic chondrogenic ATDC5 cells.

METHODS: Leptin receptor expression and iNOS messenger RNA expression were evaluated by reverse transcriptase-polymerase chain reaction. Then, iNOS activity was indirectly studied by measuring nitrite accumulation, using the Griess colorimetric reaction, in culture medium of human primary chondrocytes and ATDC5 cells.

RESULTS: ATDC5 mouse embryonic cells expressed functional OB receptor. Alone, neither OB nor IFNgamma produced nitrite accumulation in culture medium. However, costimulation with OB and IFNgamma resulted in dose-dependent up-regulation of the expression of iNOS and NO production in human primary chondrocytes and ATDC5 cells. Production of NO was blunted by the iNOS-specific inhibitors L-N(G)-nitroarginine methyl ester and aminoguanidine. In addition, the janus-activated kinase 2 (JAK2)-specific inhibitor Tyrphostin AG 490 completely blocked OB + IFNgamma-driven up-regulation of iNOS and NO production.

CONCLUSION: Our data show for the first time a putative proinflammatory role of OB via iNOS induction and NO production. This occurs via activation of JAK2.

DOI10.1002/art.10811
Alternate JournalArthritis Rheum.
PubMed ID12571850