Inhibition of furin-like enzymes blocks interleukin-1alpha/oncostatin M-stimulated cartilage degradation

Arthritis Rheum. 2003 Apr;48(4):1057-66. doi: 10.1002/art.10873.

Abstract

Objective: To investigate the role of furin-like enzymes in the proteolytic cascades leading to cartilage breakdown and to examine which collagenase(s) contribute to collagen degradation.

Methods: Bovine nasal cartilage was stimulated to resorb with the addition of interleukin-1alpha (IL-1alpha)/oncostatin M (OSM) in the presence or absence of a furin inhibitor, Dec-RVKR-CH(2)Cl, or selective matrix metalloproteinase 1 (MMP-1) inhibitors. Collagen and proteoglycan levels were determined by assay of hydroxyproline and sulfated glycosaminoglycan, respectively. Collagenase and gelatinase activity were measured using (3)H-acetylated collagen and gelatin zymography, respectively.

Results: The addition of Dec-RVKR-CH(2)Cl to stimulated cartilage reduced the release of collagen fragments and the levels of active collagenase and MMP-2, suggesting that furin-like enzymes are involved in the cascades leading to activation of procollagenases. At MMP inhibitor concentrations that selectively inhibit MMP-1, no inhibition of collagen release was observed, but increasing the concentration to the 50% inhibition concentration for MMP-13 resulted in a 50% blockage of collagen release. The addition of Dec-RVKR-CH(2)Cl to resorbing cartilage also partially blocked proteoglycan release, thus demonstrating a role for furin-activated enzymes in the pathways leading to proteoglycan degradation.

Conclusion: Furin-like enzymes are involved in cascades leading to activation of procollagenases and degradation of collagen. MMP-13, which can be activated by furin-processed membrane-type 1 MMP-1, appears to be a major collagenase involved in collagen degradation induced by IL-1alpha/OSM. Furin-like enzymes also appear to play a role in the pathways leading to proteoglycan degradation. These findings are of importance when considering proteinase inhibition as a target for therapeutic intervention in arthritic diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Chloromethyl Ketones / pharmacology
  • Animals
  • Cattle
  • Collagen / metabolism
  • Dose-Response Relationship, Drug
  • Drug Combinations
  • Enzyme Inhibitors / pharmacology*
  • Furin
  • Hydroxyproline / metabolism
  • Interleukin-1 / pharmacology*
  • L-Lactate Dehydrogenase / metabolism
  • Matrix Metalloproteinase 13
  • Matrix Metalloproteinase Inhibitors
  • Nasal Septum / drug effects*
  • Nasal Septum / enzymology
  • Oligopeptides / pharmacology*
  • Oncostatin M
  • Organ Culture Techniques
  • Peptides / pharmacology*
  • Proteoglycans / metabolism
  • Recombinant Proteins
  • Subtilisins / antagonists & inhibitors*

Substances

  • Amino Acid Chloromethyl Ketones
  • Drug Combinations
  • Enzyme Inhibitors
  • Interleukin-1
  • Matrix Metalloproteinase Inhibitors
  • Oligopeptides
  • Peptides
  • Proteoglycans
  • Recombinant Proteins
  • decanoylRVKRchloromethylketone
  • Oncostatin M
  • Collagen
  • L-Lactate Dehydrogenase
  • Subtilisins
  • Furin
  • Matrix Metalloproteinase 13
  • Hydroxyproline