Production of MMPs in human cerebral endothelial cells and their role in shedding adhesion molecules

J Neuropathol Exp Neurol. 2001 Apr;60(4):320-7. doi: 10.1093/jnen/60.4.320.

Abstract

Matrix metalloproteinases (MMPs) are Zn2+-endopeptidases that seem to play an important role in chronic inflammatory diseases of the central nervous system by disrupting the blood-brain barrier (BBB) and mediating the destruction of myelin components. We therefore investigated the influence of the pro-inflammatory cytokine TNF-alpha. on the expression and activation of several MMPs in human cerebral endothelial cells (HCEC). HCEC constitutively express MMP-2 and MMP-3 mRNA, but only MMP-3 is upregulated on mRNA and protein level after TNF-alpha stimulation. MMP-9 and MMP-12 mRNA could only be detected under inflammatory conditions. Furthermore, MMPs are involved in shedding of cell surface molecules. We therefore investigated the influence of MMPs on the release of soluble adhesion molecules using marimastat, a specific broad-spectrum MMP inhibitor and other protease inhibitors like aprotinin or leupeptin. Only marimastat inhibited the TNF-alpha mediated release of sVCAM-1 in the supernatants of HCEC. Western blot results of culture supernatants supported the time dependent release of the complete extracellular portion of the VCAM-1 molecule. These data suggest that MMPs produced by HCEC are actively involved in the shedding of soluble adhesion molecules at the BBB.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing
  • Blotting, Northern
  • Cell Adhesion Molecules / metabolism*
  • Cells, Cultured
  • Cerebral Cortex / blood supply*
  • Cerebral Cortex / cytology
  • Culture Media, Conditioned / metabolism
  • DNA-Binding Proteins
  • Dose-Response Relationship, Drug
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism*
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Gene Deletion
  • Humans
  • Matrix Metalloproteinases / genetics
  • Matrix Metalloproteinases / metabolism*
  • Microcirculation / cytology
  • Protease Inhibitors / pharmacology
  • RNA, Messenger / biosynthesis
  • Transcription Factors
  • Tumor Necrosis Factor-alpha / pharmacology
  • Vascular Cell Adhesion Molecule-1 / genetics
  • Vascular Cell Adhesion Molecule-1 / metabolism

Substances

  • Cell Adhesion Molecules
  • Culture Media, Conditioned
  • DNA-Binding Proteins
  • Protease Inhibitors
  • RNA, Messenger
  • Transcription Factors
  • Tumor Necrosis Factor-alpha
  • Vascular Cell Adhesion Molecule-1
  • Matrix Metalloproteinases