Inducible hydrogen sulfide synthesis in chondrocytes and mesenchymal progenitor cells: is H2S a novel cytoprotective mediator in the inflamed joint?
Bridget Fox
Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK
Search for more papers by this authorJan-Thorsten Schantz
Department of Plastic, Reconstructive and Handsurgery, Klinikum rechts der Isar, Technische Universität München, München, Germany
Division of Plastic Surgery and Bioengineering, National University of Singapore, Singapore
Search for more papers by this authorRichard Haigh
Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK
Department of Rheumatology, Royal Devon and Exeter Hospital Trust, Exeter, Devon, UK
Search for more papers by this authorMark E. Wood
Biosciences, College of Life and Environmental Sciences of Biosciences, University of Exeter, Streatham Campus, Exeter, Devon, UK
Search for more papers by this authorPhillip K. Moore
Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
Search for more papers by this authorNick Viner
Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK
Department of Rheumatology, Torbay Hospital, Torbay, Devon, UK
Search for more papers by this authorJeremy P. E. Spencer
Department of Chemistry and Food Biosciences, University of Reading, Whiteknights, Berkshire, UK
Search for more papers by this authorPaul G. Winyard
Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK
Search for more papers by this authorCorresponding Author
Matthew Whiteman
Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
Matthew WHITEMAN, Peninsula Medical School, University of Exeter, St. Luke’s Campus, Magdalen Road, Exeter, Devon EX1 2LU, UK. Tel.:+44(0)1392-722942 E-mail: [email protected] or [email protected]Search for more papers by this authorBridget Fox
Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK
Search for more papers by this authorJan-Thorsten Schantz
Department of Plastic, Reconstructive and Handsurgery, Klinikum rechts der Isar, Technische Universität München, München, Germany
Division of Plastic Surgery and Bioengineering, National University of Singapore, Singapore
Search for more papers by this authorRichard Haigh
Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK
Department of Rheumatology, Royal Devon and Exeter Hospital Trust, Exeter, Devon, UK
Search for more papers by this authorMark E. Wood
Biosciences, College of Life and Environmental Sciences of Biosciences, University of Exeter, Streatham Campus, Exeter, Devon, UK
Search for more papers by this authorPhillip K. Moore
Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
Search for more papers by this authorNick Viner
Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK
Department of Rheumatology, Torbay Hospital, Torbay, Devon, UK
Search for more papers by this authorJeremy P. E. Spencer
Department of Chemistry and Food Biosciences, University of Reading, Whiteknights, Berkshire, UK
Search for more papers by this authorPaul G. Winyard
Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK
Search for more papers by this authorCorresponding Author
Matthew Whiteman
Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
Matthew WHITEMAN, Peninsula Medical School, University of Exeter, St. Luke’s Campus, Magdalen Road, Exeter, Devon EX1 2LU, UK. Tel.:+44(0)1392-722942 E-mail: [email protected] or [email protected]Search for more papers by this authorAbstract
Hydrogen sulfide (H2S) has recently been proposed as an endogenous mediator of inflammation and is present in human synovial fluid. This study determined whether primary human articular chondrocytes (HACs) and mesenchymal progenitor cells (MPCs) could synthesize H2S in response to pro-inflammatory cytokines relevant to human arthropathies, and to determine the cellular responses to endogenous and pharmacological H2S. HACs and MPCs were exposed to IL-1β, IL-6, TNF-α and lipopolysaccharide (LPS). The expression and enzymatic activity of the H2S synthesizing enzymes cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE) were determined by Western blot and zinc-trap spectrophotometry, respectively. Cellular oxidative stress was induced by H2O2, the peroxynitrite donor SIN-1 and 4-hydroxynonenal (4-HNE). Cell death was assessed by 3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. Mitochondrial membrane potential (DCm) was determined in situ by flow cytometry. Endogenous H2S synthesis was inhibited by siRNA-mediated knockdown of CSE and CBS and pharmacological inhibitors D,L-propargylglycine and aminoxyacetate, respectively. Exogenous H2S was generated using GYY4137. Under basal conditions HACs and MPCs expressed CBS and CSE and synthesized H2S in a CBS-dependent manner, whereas CSE expression and activity was induced by treatment of cells with IL-1β, TNF-α, IL-6 or LPS. Oxidative stress-induced cell death was significantly inhibited by GYY4137 treatment but increased by pharmacological inhibition of H2S synthesis or by CBS/CSE-siRNA treatment. These data suggest CSE is an inducible source of H2S in cultured HACs and MPCs. H2S may represent a novel endogenous mechanism of cytoprotection in the inflamed joint, suggesting a potential opportunity for therapeutic intervention.
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