Destruction of articular cartilage by alpha 2 macroglobulin elastase complexes: role in rheumatoid arthritis.
OBJECTIVE
Neutrophil elastase accounts for the ability of some fresh rheumatoid synovial fluids to degrade cartilage matrix in vitro. The aim of this study was to determine if enzyme activity could result from depletion of synovial fluid inhibitors or protection of the enzyme from inhibition.


METHODS
The ability of synovial fluids to inhibit porcine pancreatic elastase was investigated together with chemical pretreatments capable of inactivating alpha 1 protease inhibitor (alpha 1PI) or preventing formation of alpha 2 macroglobulin (alpha 2M) elastase complexes. Subsequently, complexes of human neutrophil elastase with alpha 2M were prepared and applied to frozen sections of cartilage. Proteoglycan loss was quantified by alcian blue staining and scanning and integrating microdensitometry. Parallel studies were carried out using a low molecular weight chromogenic elastase substrate. The effects of alpha 1PI and SF on these systems were investigated. Finally, synovial fluids were subjected to gel filtration and the fractions assayed for elastase activity. High molecular weight fractions were pooled, concentrated, and tested for their ability to degrade cartilage sections.


RESULTS
All synovial fluids reduced the activity of porcine pancreatic elastase, the inhibition mainly being attributable to alpha 1PI, whereas remaining activity resulted from complexes of elastase with alpha 2M. Complexes of human neutrophil elastase with alpha 2M were shown to cause proteoglycan degradation in frozen sections of human articular cartilage. Alpha 1PI prevented alpha 2M elastase complexes from degrading cartilage but not the chromogenic substrate. The data suggested that alpha 1PI does not inhibit elastase bound to alpha 2M but sterically hinders the complex. However, only one of five synovial fluids was able to completely block the actions of alpha 2M elastase complexes against cartilage. Gel filtration of rheumatoid synovial fluids showed elastase and cartilage degrading activity to be associated with fractions that contained alpha 2M, and not with fractions expected to contain free enzyme.


CONCLUSIONS
The data suggest that synovial fluid alpha 2M elastase complexes can degrade cartilage matrix in rheumatoid arthritis.
Moore
A R
AR
Department of Experimental Pathology, St Bartholomew's, London.
Appelboam
A
A
Kawabata
K
K
Da Silva
J A
JA
D'Cruz
D
D
Gowland
G
G
Willoughby
D A
DA
D000328:Adult; D001172:Arthritis, Rheumatoid / Q000378:metabolism*; D002358:Cartilage, Articular / Q000378:metabolism*; D002845:Chromatography; D002850:Chromatography, Gel; D006801:Humans; D007719:Knee Joint; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011485:Protein Binding; D011509:Proteoglycans / Q000378:metabolism; D013582:Synovial Fluid / Q000201:enzymology / Q000378:metabolism*; D000515:alpha 1-Antitrypsin / Q000378:metabolism; D000511:alpha-Macroglobulins / Q000378:metabolism*
D011509:Proteoglycans; D000515:alpha 1-Antitrypsin; D000511:alpha-Macroglobulins; D019272:Leukocyte Elastase