Cysteine protease activity is up-regulated in inflamed ankle joints of rats with adjuvant-induced arthritis and decreases with in vivo administrationof a vinyl sulfone cysteine protease inhibitor
Sl. Biroc et al., Cysteine protease activity is up-regulated in inflamed ankle joints of rats with adjuvant-induced arthritis and decreases with in vivo administrationof a vinyl sulfone cysteine protease inhibitor, ARTH RHEUM, 44(3), 2001, pp. 703-711
Objective. Cysteine proteases are postulated to play a role in tissue destr
uction in the joints of animals with arthritis. The purpose of the present
study was to confirm the concept that cysteine proteases are enzymes involv
ed in the pathology of rheumatoid arthritis (RA).
Methods. Arthritis was induced in Lewis rats by adjuvant injection (adjuvan
t-induced arthritis [AIA] model) and scored for inflammation. At necropsy,
the rear paws were either fixed in formalin and assigned a histologic score
(based on synovial cell proliferation, cartilage erosion, bone erosion, an
d fibroproliferative pannus) or frozen, cryosectioned, and assayed for enzy
me activity either by in situ cytochemical staining with a post-azo-couplin
g method using a chromogenic substrate (Z-arg-arg-MNA) or by a novel assay
placing the tissue section directly in a cuvette using the fluorogenic subs
trate Z-arg-arg-AMC.
Results. Enzymatic activity, measured either in frozen sections in situ or
in the cuvette assay, was positively correlated with joint destruction (r =
0.7) and inflammation (r = 0.8). Activity was not inhibited significantly
by Pefabloc (a serine protease inhibitor), EDTA (a metalloprotease inhibito
r), or pepstatin A (an aspartyl protease inhibitor) but was inhibited by E-
64 and vinyl sulfone irreversible inhibitors of cysteine proteases. The eff
ect of one of the vinyl sulfone cysteine protease inhibitors, Mu-Leu-HomoPh
e-vinylsulfone, was tested in vivo by dietary administration at 2.2 mg/kg/d
ay in the AIA model; this resulted in a significant decrease in inflammatio
n and in the amount of cysteine protease activity measured in the joint tis
sue.
Conclusion. Cysteine protease activity levels increase in the diseased stat
e and may be an important target for designing small molecule inhibitors to
reduce the inflammation and tissue destruction associated with RA.