Hydrogen sulfide (H(2) S) 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 H(2) S in response to pro-inflammatory cytokines relevant to human arthropathies, and to determine the cellular responses to endogenous and pharmacological H(2) S. HACs and MPCs were exposed to IL-1?, IL-6, TNF-? and lipopolysaccharide (LPS). The expression and enzymatic activity of the H(2) S synthesizing enzymes cystathionine-?-synthase (CBS) and cystathionine-?-lyase (CSE) were determined by Western blot and zinc-trap spectrophotometry, respectively. Cellular oxidative stress was induced by H(2) O(2) , 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 H(2) S synthesis was inhibited by siRNA-mediated knockdown of CSE and CBS and pharmacological inhibitors D,L-propargylglycine and aminoxyacetate, respectively. Exogenous H(2) S was generated using GYY4137. Under basal conditions HACs and MPCs expressed CBS and CSE and synthesized H(2) S in a CBS-dependent manner, whereas CSE expression and activity was induced by treatment of cells with IL-1?, TNF-?, IL-6 or LPS. Oxidative stress-induced cell death was significantly inhibited by GYY4137 treatment but increased by pharmacological inhibition of H(2) S synthesis or by CBS/CSE-siRNA treatment. These data suggest CSE is an inducible source of H(2) S in cultured HACs and MPCs. H(2) S may represent a novel endogenous mechanism of cytoprotection in the inflamed joint, suggesting a potential opportunity for therapeutic intervention.     «

Hydrogen sulfide (H(2) S) 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 H(2) S in response to pro-inflammatory cytokines relevant to human arthropathies, and to determine the cellular responses to endogenous and pharmacological H(2) S. HACs and MPCs were exposed to IL-1?, IL-6, TNF-? and lipopolysaccharid...     »