Inducible hydrogen sulfide synthesis in chondrocytes and mesenchymal progenitor cells: is H2S a novel cytoprotective mediator in the inflamed joint?

Journal of Cellular and Molecular Medicine

Bridget Fox⁵  Jan-Thorsten Schantz²  Richard Haigh⁵  Mark E. Wood⁴  Phillip K. Moore³  Nick Viner⁵  Jeremy P. E. Spencer¹  Paul G. Winyard⁵ 

[1] Department of Chemistry and Food Biosciences, University of Reading, Whiteknights, Berkshire, UK;Department of Plastic, Reconstructive and Handsurgery, Klinikum rechts der Isar, Technische Universität München, München, Germany;Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore;Biosciences, College of Life and Environmental Sciences of Biosciences, University of Exeter, Streatham Campus, Exeter, Devon, UK;Peninsula Medical School, University of Exeter, St. Luke’s Campus, Exeter, Devon, UK

关键词: arthritis;    cystathionine‐γ‐lyase;    cystathionine‐β‐synthase;    GYY4137;    apoptosis;    oxidative stress;   

DOI  :  10.1111/j.1582-4934.2011.01357.x

来源: Wiley

【 摘 要 】

Abstract

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.

【 授权许可】

Unknown   
© 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd