Measurement of elastase and cysteine proteinases in synovial fluid of patients with rheumatoid arthritis, sero-negative spondylarthropathies, and osteoarthritis.
Synovial fluid samples were collected from 45 patients with rheumatoid arthritis, spondylarthropathy, or osteoarthritis, to study their content of elastase (EC 3.4.21.37) and of cysteine proteinases (EC 3.4.22.1, 3.4.22.15). We measured both elastase complexed with alpha 1-proteinase inhibitor and elastase activity toward the substrate L-pyroglutamyl-L-prolyl-L-valine-p-nitroanilide. Cysteine proteinase activities were measured with the substrates N-benzyloxycarbonyl-L-phenylalanyl-L-arginine-7-amido-4-methylcoumarin (Z-Phe-Arg-AMC) and Z-Arg-Arg-AMC and the inhibitor E-64 [L-trans-epoxysuccinyl-leucyl-amido-(4-guanidino)-butane]. In all these enzyme assays, higher median values were obtained in inflammatory arthropathies than in osteoarthritis. The concentration of the elastase-alpha 1-proteinase inhibitor complex and of elastase and cysteine proteinase activities were statistically higher in patients with rheumatoid arthritis than in patients with osteoarthritis. The difference in results between patients with spondylarthropathy and patients with osteoarthritis was statistically significant only for the elastase-alpha 1-proteinase inhibitor complex. The median values of the complex and of both enzyme activities were higher in patients with rheumatoid arthritis than in patients with spondylarthropathy; however, the difference was statistically significant only for the cysteine proteinase activity measured with Z-Arg-Arg-AMC substrate. These results suggest that both elastase and cysteine proteinases, which are increased in patients with inflammatory arthritis, are involved in cartilage degradation in these arthropathies.
Huet
G
G
Laboratoire de Biochimie, Hôpital Huriez, Lille, France.
Flipo
R M
RM
Richet
C
C
Thiebaut
C
C
Demeyer
D
D
Balduyck
M
M
Duquesnoy
B
B
Degand
P
P
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D001172:Arthritis, Rheumatoid / Q000201:enzymology*; D003374:Coumarins / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism; D004151:Dipeptides / Q000378:metabolism; D005260:Female; D006801:Humans; D007930:Leucine / Q000031:analogs & derivatives / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010003:Osteoarthritis / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000378:metabolism*; D013122:Spinal Diseases / Q000201:enzymology*; D013379:Substrate Specificity; D013582:Synovial Fluid / Q000201:enzymology*
D003374:Coumarins; D015853:Cysteine Proteinase Inhibitors; D004151:Dipeptides; C021159:benzyloxycarbonyl-phenylalanylarginine-4-methylcoumaryl-7-amide; D010196:Pancreatic Elastase; D003546:Cysteine Endopeptidases; D007930:Leucine; C024974:E 64