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dc.contributor.authorOtero, Miguel
dc.contributor.authorLago, Rocío
dc.contributor.authorGómez-Reino Carnota, Juan Jesús
dc.contributor.authorGualillo, Oreste
dc.date.accessioned2020-06-12T08:43:32Z
dc.date.available2020-06-12T08:43:32Z
dc.date.issued2005
dc.identifier.citationOtero, M., Lago, R., Lago, F. et al. Signalling pathway involved in nitric oxide synthase type II activation in chondrocytes: synergistic effect of leptin with interleukin-1. Arthritis Res Ther 7, R581 (2005). https://doi.org/10.1186/ar1708
dc.identifier.issn1478-6354
dc.identifier.urihttp://hdl.handle.net/10347/22953
dc.description.abstractThe objective of the present study was to investigate the effect of leptin, alone or in combination with IL-1, on nitric oxide synthase (NOS) type II activity in vitro in human primary chondrocytes, in the mouse chondrogenic ATDC5 cell line, and in mature and hypertrophic ATDC5 differentiated chondrocytes. For completeness, we also investigated the signalling pathway of the putative synergism between leptin and IL-1. For this purpose, nitric oxide production was evaluated using the Griess colorimetric reaction in culture medium of cells stimulated over 48 hours with leptin (800 nmol/l) and IL-1 (0.025 ng/ml), alone or combined. Specific pharmacological inhibitors of NOS type II (aminoguanidine [1 mmol/l]), janus kinase (JAK)2 (tyrphostin AG490 and Tkip), phosphatidylinositol 3-kinase (PI3K; wortmannin [1, 2.5, 5 and 10 μmol/l] and LY294002 [1, 2.5, 5 and 10 μmol/l]), mitogen-activated protein kinase kinase (MEK)1 (PD098059 [1, 5, 10, 20 and 30 μmol/l]) and p38 kinase (SB203580 [1, 5, 10, 20 and 30 μmol/l]) were added 1 hour before stimulation. Nitric oxide synthase type II mRNA expression in ATDC5 chondrocytes was investigated by real-time PCR and NOS II protein expression was analyzed by western blot. Our results indicate that stimulation of chondrocytes with IL-1 results in dose-dependent nitric oxide production. In contrast, leptin alone was unable to induce nitric oxide production or expression of NOS type II mRNA or its protein. However, co-stimulation with leptin and IL-1 resulted in a net increase in nitric oxide concentration over IL-1 challenge that was eliminated by pretreatment with the NOS II specific inhibitor aminoguanidine. Pretreatment with tyrphostin AG490 and Tkip (a SOCS-1 mimetic peptide that inhibits JAK2) blocked nitric oxide production induced by leptin/IL-1. Finally, wortmannin, LY294002, PD098059 and SB203580 significantly decreased nitric oxide production. These findings were confirmed in mature and hypertrophic ATDC5 chondrocytes, and in human primary chondrocytes. This study indicates that leptin plays a proinflammatory role, in synergy with IL-1, by inducing NOS type II through a signalling pathway that involves JAK2, PI3K, MEK-1 and p38 kinase
dc.description.sponsorshipThis work was supported by grants from Spanish Ministry of Health (FIS 01/3137 and PI-020431). Oreste Gualillo and Francisca Lago are recipients of a research contract from Spanish Ministry of Health, Instituto de Salud Carlos III (EXP 00/3051 and 99/3040). Miguel Otero is a recipient of a predoctoral fellowship funded by Xunta de Galicia. Rocío Lago is a recipient of a fellowship funded by Instituto de Salud Carlos III (Red Temática G03/152)
dc.language.isoeng
dc.publisherBMC
dc.rights© 2005 Otero et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/ 2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited
dc.rights.urihttp://creativecommons.org/licenses/by/2.0
dc.subjectNitric Oxide
dc.subjectSB203580
dc.subjectWortmannin
dc.subjectHuman Chondrocytes
dc.subjectNitrite Accumulation
dc.titleSignalling pathway involved in nitric oxide synthase type II activation in chondrocytes: synergistic effect of leptin with interleukin-1
dc.typeinfo:eu-repo/semantics/article
dc.identifier.DOI10.1186/ar1708
dc.relation.publisherversionhttps://doi.org/10.1186/ar1708
dc.type.versioninfo:eu-repo/semantics/publishedVersion
dc.identifier.e-issn1478-6362
dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.contributor.affiliationUniversidade de Santiago de Compostela. Departamento de Psiquiatría, Radioloxía, Saúde Pública, Enfermaría e Medicina
dc.description.peerreviewedSI


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© 2005 Otero et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/ 2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited
Except where otherwise noted, this item's license is described as  © 2005 Otero et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/ 2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited





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