Synergistic Effect of Thrombin and CD40 Ligand on Endothelial Matrix metalloproteinase-10 Expression and Microparticle Generation in Vitro and in Vivo

Arterioscler Thromb Vasc Biol. 2012 Jun;32(6):1477-87. doi: 10.1161/ATVBAHA.112.248773. Epub 2012 Apr 5.

Abstract

Objective: Thrombin induces CD40 ligand (CD40L) and matrix metalloproteinases (MMPs) under inflammatory/prothrombotic conditions. Thrombin and CD40L could modulate endothelial MMP-10 expression in vitro and in vivo.

Methods and results: Human endothelial cells were stimulated with thrombin (0.1-10 U/mL), CD40L (0.25-1 μg/mL), or their combination (thrombin/CD40L) to assess MMP-10 expression and microparticle generation. Thrombin/CD40L elicited higher MMP-10 mRNA (5-fold; P<0.001) and protein levels (4.5-fold; P<0.001) than either stimulus alone. This effect was mimicked by a protease-activated receptor-1 agonist and antagonized by hirudin, a-protease-activated receptor-1, α-CD40L, and α-CD40 antibodies. The synergistic effect was dependent on p38 mitogen-activated protein kinase and c-Jun N-terminal kinase-1 pathways. Thrombin also upregulated the expression of CD40 in endothelial cell surface increasing its availability, thereby favoring its synergistic effects with CD40L. In mice, thrombin/CD40L further increased the aortic MMP-10 expression. Septic patients with systemic inflammation and enhanced thrombin generation (n=60) exhibited increased MMP-10 and soluble CD40L levels associated with adverse clinical outcome. Endothelial and systemic activation by thrombin/CD40L and lipopolysaccharide also increased microparticles harboring MMP-10 and CD40L.

Conclusions: Thrombin/CD40L elicited a strong synergistic effect on endothelial MMP-10 expression and microparticles containing MMP-10 in vitro and in vivo, which may represent a new link between inflammation/thrombosis with prognostic implications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Animals
  • Antibodies / pharmacology
  • Blood Coagulation
  • CD40 Antigens / antagonists & inhibitors
  • CD40 Antigens / metabolism
  • CD40 Ligand / antagonists & inhibitors
  • CD40 Ligand / blood
  • CD40 Ligand / metabolism*
  • Case-Control Studies
  • Cell-Derived Microparticles / enzymology*
  • Cell-Derived Microparticles / pathology
  • Cells, Cultured
  • Disease Models, Animal
  • Disseminated Intravascular Coagulation / enzymology
  • Disseminated Intravascular Coagulation / pathology
  • Endothelial Cells / drug effects
  • Endothelial Cells / enzymology*
  • Endothelial Cells / pathology
  • Endotoxemia / enzymology
  • Endotoxemia / genetics
  • Endotoxemia / pathology
  • Female
  • Gene Expression Regulation, Enzymologic
  • Hirudins / pharmacology
  • Human Umbilical Vein Endothelial Cells / enzymology
  • Humans
  • Lipopolysaccharides / pharmacology
  • Male
  • Matrix Metalloproteinase 10 / blood
  • Matrix Metalloproteinase 10 / deficiency
  • Matrix Metalloproteinase 10 / genetics
  • Matrix Metalloproteinase 10 / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Middle Aged
  • Mitogen-Activated Protein Kinase 8 / antagonists & inhibitors
  • Mitogen-Activated Protein Kinase 8 / metabolism
  • Multivariate Analysis
  • Peptides / pharmacology
  • Protein Kinase Inhibitors / pharmacology
  • RNA, Messenger / metabolism
  • Receptor, PAR-1 / agonists
  • Receptor, PAR-1 / antagonists & inhibitors
  • Receptor, PAR-1 / metabolism
  • Risk Assessment
  • Risk Factors
  • Sepsis / enzymology*
  • Sepsis / mortality
  • Sepsis / pathology
  • Signal Transduction
  • Spain
  • Survival Analysis
  • Thrombin / metabolism*
  • Time Factors
  • Up-Regulation
  • p38 Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • Antibodies
  • CD40 Antigens
  • Hirudins
  • Lipopolysaccharides
  • Peptides
  • Protein Kinase Inhibitors
  • RNA, Messenger
  • Receptor, PAR-1
  • CD40 Ligand
  • Mitogen-Activated Protein Kinase 8
  • p38 Mitogen-Activated Protein Kinases
  • Thrombin
  • MMP10 protein, human
  • Matrix Metalloproteinase 10