Mechanisms of human neutrophil-mediated cartilage damage in vitro: the role of lysosomal enzymes, hydrogen peroxide and hypochlorous acid.
Cartilage is a focal point of attack by cellular and molecular elements of the inflammatory response which occurs in arthritic diseases. Neutrophils damage articular cartilage by degrading matrix components and inhibiting their synthesis. The aim of this study was to elucidate mechanisms of this damage. Human neutrophils were isolated from blood by centrifuging through Ficoll-Hypaque and granule extract prepared from them. Articular cartilage from adult humans and cattle was maintained in organ culture. Cartilage degradation (release of 35S-labelled proteoglycan) or synthesis (incorporation of 35S into proteoglycan) was determined after various treatments. Human neutrophils and neutrophil granule extract degraded proteoglycan and inhibited proteoglycan synthesis. The specific leucocyte elastase inhibitor N-methoxysuccinyl-(ala)2-pro-val-chloromethylketone (MAAPVCMK) partially reversed these effects. H2O2, a product of the neutrophil respiratory burst, when added directly at 10(-6)mol/L, or generated by glucose oxidase (GO)/glucose inhibited proteoglycan synthesis but had no effect on degradation. Hypochlorous acid (OHCl), a product of the myeloperoxidase (MPO)/H2O2/Cl system at 50 mumol/L degraded proteoglycan and inhibited its synthesis. OHCl produced by granule extract (as a source of MPO) + GO-generated H2O2 + Cl- degraded proteoglycan. The results indicate that neutrophil-mediated proteoglycan degradation and inhibition of synthesis is largely attributable to elastase and secondarily to OHCl, whereas H2O2 impairs synthesis without affecting degradation of proteoglycan.
Kowanko
I C
IC
Department of Immunology, Adelaide Children's Hospital, Australia.
Bates
E J
EJ
Ferrante
A
A
D000818:Animals; D002357:Cartilage Diseases / Q000201:enzymology / Q000209:etiology* / Q000276:immunology; D002358:Cartilage, Articular; D002417:Cattle; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D006997:Hypochlorous Acid / Q000378:metabolism*; D008247:Lysosomes / Q000201:enzymology*; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism*; D009924:Organ Culture Techniques; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism
D011509:Proteoglycans; D006997:Hypochlorous Acid; D006861:Hydrogen Peroxide; D010196:Pancreatic Elastase