Partial purification of gelatinases and effect of anti-inflammatory drugs on the tumor necrosis factor-stimulated production of active gelatinase by granulation tissue in culture.
Gelatinases produced by granulation tissue in culture were separated into two fractions when the conditioned medium was chromatographed on diethylaminoethyl-Sephacel; one contained latent and active gelatinases with molecular weight of about 74 kDa (low-Mr gelatinase), and the other contained an active gelatinase with molecular weight of about 112 kDa (high-Mr gelatinase) as estimated by gel filtration on Sephadex G-150. The former was unbound and the latter was bound to Zn-chelating Sepharose. Both low- and high-Mr gelatinases, however, occurred in mainly two bands corresponding to molecular weights of 64 and 57 kDa on sodium dodecyl sulfate-substrate polyacrylamide gel electrophoresis. Recombinant human tumor necrosis factor-alpha (TNF) markedly enhanced the production of gelatinases, especially high-Mr active gelatinase. Dexamethasone and hydrocortisone strongly suppressed TNF-mediated production of high-Mr active gelatinase, whereas indomethacin, piroxicam and mepacrine had no effect. The results suggest that steroidal anti-inflammatory drugs suppress a rapid collagen breakdown in granulation tissue through their inhibitory actions--one of which is the suppression of active gelatinase production enhanced by cytokines including TNF.
Nakagawa
H
H
Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Japan.
Aikawa
Y
Y
Kitagawa
H
H
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D046508:Culture Techniques; D003907:Dexamethasone / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D018093:Gelatinases; D006097:Granulation Tissue / Q000187:drug effects* / Q000201:enzymology; D006854:Hydrocortisone / Q000494:pharmacology; D008297:Male; D010434:Pepsin A / Q000096:biosynthesis / Q000302:isolation & purification*; D051381:Rats; D011919:Rats, Inbred Strains; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D000893:Anti-Inflammatory Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone; D010434:Pepsin A; D018093:Gelatinases; D006854:Hydrocortisone