PMID- 26093672 OWN - NLM STAT- MEDLINE DCOM- 20160204 LR - 20190816 IS - 1471-2474 (Electronic) IS - 1471-2474 (Linking) VI - 16 DP - 2015 Jun 21 TI - alpha-Melanocyte-stimulating-hormone (alpha-MSH) modulates human chondrocyte activation induced by proinflammatory cytokines. PG - 154 LID - 10.1186/s12891-015-0615-1 [doi] LID - 154 AB - BACKGROUND: Alpha-melanocyte-stimulating-hormone (alpha-MSH) has marked anti-inflammatory potential. Proinflammatory cytokines are critical mediators of the disturbed cartilage homeostasis in osteoarthritis, inhibiting anabolic activities and increasing catabolic activities in chondrocytes. Since human chondrocytes express alpha-MSH receptors, we evaluated the role of the peptide in modulating chondrocyte production of pro-inflammatory cytokines, matrix metalloproteinases (MMPs), tissue inhibitors of MMPs (TIMPs), inducible nitric oxide synthase (iNOS) and nitric oxide (NO) in response to interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha). METHODS: Human articular chondrocytes were obtained from osteoarthritic joint cartilage from subjects undergoing hip routine arthroplasty procedures. The cells were cultured with or without alpha-MSH in the presence of IL-1beta or TNF-alpha. Cell-free supernatants were collected and cells immediately lysed for RNA purification. Expression of cytokines, MMPs, TIMPs, iNOS was determined by Reverse Transcription Real-time Polymerase Chain Reaction and enzyme-linked immunosorbent assay. Griess reaction was used for NO quantification. RESULTS: Gene expression and secretion of IL-6, IL-8, MMP-3, MMP-13 were significantly increased in IL-1beta or TNF-alpha-stimulated chondrocytes; alpha-MSH did not modify the release of IL-6 or IL-8 while the peptide significantly reduced their gene expression on TNF-alpha-stimulated cells. A significant inhibition of MMP3 gene expression and secretion from IL-1beta or TNFalpha-stimulated chondrocytes was induced by alpha-MSH. On the other hand, alpha-MSH did not modify the release of MMP-13 by cytokine-stimulated chondrocyte but significantly decreased gene expression of the molecule on TNF-alpha-stimulated cells. Detectable amount of TIMP-3 and TIMP-4 were present in the supernatants of resting chondrocytes and a significant increase of TIMP-3 gene expression and release was induced by alpha-MSH on unstimulated cells. TIMP-3 secretion and gene expression were significantly increased in IL-1beta-stimulated chondrocytes and alpha-MSH down-regulated gene expression but not secretion of the molecule. TIMP-4 gene expression (but not secretion) was moderately induced in IL-1beta-stimulated chondrocytes with a down-regulation exerted by alpha-MSH. IL-1beta and TNF-alpha were potent stimuli for NO production and iNOS gene expression by chondrocytes; no inhibition was induced by alpha-MSH on cytokine-stimulated NO production, while the peptide significantly reduced gene expression of iNOS. CONCLUSIONS: Our results underscore a potential anti-inflammatory and chondroprotective activity exerted by alpha-MSH, increasing TIMP-3 gene expression and release on resting cells and down- modulating TNF-alpha-induced activation of human chondrocytes. However, the discrepancy between the influences exerted by alpha-MSH on gene expression and protein release as well as the difference in the inhibitory pattern exerted by alpha-MSH in TNF-alpha- or IL-1beta-stimulated cells leave some uncertainty on the role of the peptide on chondrocyte modulation. FAU - Capsoni, Franco AU - Capsoni F AD - Allergy, Clinical Immunology & Rheumatology Unit, Istituto Auxologico Italiano, IRCCS, University of Milan, Piazzale Brescia, 20 - 20149, Milano, Italy. franco.capsoni@unimi.it. FAU - Ongari, Anna Maria AU - Ongari AM AD - Allergy, Clinical Immunology & Rheumatology Unit, Istituto Auxologico Italiano, IRCCS, University of Milan, Piazzale Brescia, 20 - 20149, Milano, Italy. anna.ongari@unimi.it. FAU - Lonati, Caterina AU - Lonati C AD - Center for Preclinical Investigation, Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena, Milan, Italy. caterina.lonati@libero.it. FAU - Accetta, Riccardo AU - Accetta R AD - Traumatology and First Aid Unit, Istituto Ortopedico Galeazzi, IRCCS, Milan, Italy. riccardo.accetta@grupposandonato.it. FAU - Gatti, Stefano AU - Gatti S AD - Center for Preclinical Investigation, Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena, Milan, Italy. stefano.gatti@unimi.it. FAU - Catania, Anna AU - Catania A AD - Center for Preclinical Investigation, Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena, Milan, Italy. annapia.catania@guest.unimi.it. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150621 PL - England TA - BMC Musculoskelet Disord JT - BMC musculoskeletal disorders JID - 100968565 RN - 0 (Anti-Inflammatory Agents) RN - 0 (CXCL8 protein, human) RN - 0 (IL6 protein, human) RN - 0 (Inflammation Mediators) RN - 0 (Interleukin-1beta) RN - 0 (Interleukin-6) RN - 0 (Interleukin-8) RN - 0 (Tissue Inhibitor of Metalloproteinases) RN - 0 (Tumor Necrosis Factor-alpha) RN - 31C4KY9ESH (Nitric Oxide) RN - 581-05-5 (alpha-MSH) RN - 75921-69-6 (MSH, 4-Nle-7-Phe-alpha-) RN - EC 1.14.13.39 (NOS2 protein, human) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type II) RN - EC 3.4.24.- (Matrix Metalloproteinases) SB - IM MH - Anti-Inflammatory Agents/*pharmacology MH - Cells, Cultured MH - Chondrocytes/*drug effects/immunology/metabolism MH - Gene Expression Regulation MH - Humans MH - Inflammation Mediators/*pharmacology MH - Interleukin-1beta/*pharmacology MH - Interleukin-6/genetics/metabolism MH - Interleukin-8/genetics/metabolism MH - Matrix Metalloproteinases/genetics/metabolism MH - Nitric Oxide/metabolism MH - Nitric Oxide Synthase Type II/genetics/metabolism MH - Osteoarthritis, Hip/genetics/immunology/*metabolism MH - Tissue Inhibitor of Metalloproteinases/genetics/metabolism MH - Tumor Necrosis Factor-alpha/*pharmacology MH - alpha-MSH/*analogs & derivatives/pharmacology PMC - PMC4475285 EDAT- 2015/06/22 06:00 MHDA- 2016/02/05 06:00 PMCR- 2015/06/21 CRDT- 2015/06/22 06:00 PHST- 2015/02/24 00:00 [received] PHST- 2015/06/08 00:00 [accepted] PHST- 2015/06/22 06:00 [entrez] PHST- 2015/06/22 06:00 [pubmed] PHST- 2016/02/05 06:00 [medline] PHST- 2015/06/21 00:00 [pmc-release] AID - 10.1186/s12891-015-0615-1 [pii] AID - 615 [pii] AID - 10.1186/s12891-015-0615-1 [doi] PST - epublish SO - BMC Musculoskelet Disord. 2015 Jun 21;16:154. doi: 10.1186/s12891-015-0615-1.