Potential roles of hypochlorous acid and N-chloroamines in collagen breakdown by phagocytic cells in synovitis.
We have tested the effects of the neutrophil/macrophage products, hypochlorous acid (HOCl) and N-chloroamines, on the structural integrity and proteolytic susceptibility of collagen to determine if these agents could play a role in inflammatory joint destruction. Rates of HOCl reaction with collagen, and collagen gelation were monitored by spectrophotometric methods. Direct fragmentation, and degradation by collagenase were measured by the release of acid-soluble counts from 3H-collagen. Physiologically relevant concentrations of HOCl (5-50 microM) reacted rapidly and quantitatively at several sites in the collagen polypeptide chain, causing extensive protein fragmentation and preventing collagen gelation. In contrast, reaction with (5-50 microM) N-chloroalanine induced little direct collagen fragmentation. Oxidative damage by N-chloroamines was, however, evident because collagen displayed greatly increased proteolytic susceptibility following N-chloroamine treatment. Collagen degradation by collagenase increased as much as 3-fold after exposure to N-chloroamine treatment. Collagen degradation by collagenase increased as much as 3-fold after exposure to N-chloroalanine. N-chloroleucine caused a small increase in proteolytic susceptibility, but N-chlorotaurine had no effect. Collagen fragmentation by HOCl, inhibition of gelation by HOCl, and N-chloroalanine-induced proteolytic susceptibility, all increased with linear kinetics at oxidant concentrations of 5 microM to 1.0 mM. In synovitis, phagocytes expose collagen to HOCl, N-chloroamines, and collagenase. It is known that HOCl can activate neutrophil procollagenase. Based on our new findings, we propose a model of inflammatory joint destruction that also includes collagen fragmentation, and increased susceptibility of collagen to degradation by collagenase.(ABSTRACT TRUNCATED AT 250 WORDS)
Davies
J M
JM
Department of Medicine, Albany Medical College, NY 12208.
Horwitz
D A
DA
Davies
K J
KJ
D000818:Animals; D002417:Cattle; D002700:Chloramines / Q000378:metabolism* / Q000494:pharmacology; D003094:Collagen / Q000187:drug effects / Q000378:metabolism*; D017364:Collagenases / Q000378:metabolism; D005609:Free Radicals; D006997:Hypochlorous Acid / Q000378:metabolism* / Q000494:pharmacology; D066298:In Vitro Techniques; D007700:Kinetics; D008954:Models, Biological; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000378:metabolism; D010586:Phagocytes / Q000378:metabolism; D013585:Synovitis / Q000209:etiology* / Q000378:metabolism
D002700:Chloramines; D005609:Free Radicals; D010446:Peptide Fragments; D006997:Hypochlorous Acid; D003094:Collagen; D017364:Collagenases