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
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Research Support, Non-U.S. Gov't
MeSH terms
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Adult
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Aged
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Aged, 80 and over
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Animals
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Antibodies / pharmacology
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Blood Coagulation
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CD40 Antigens / antagonists & inhibitors
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CD40 Antigens / metabolism
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CD40 Ligand / antagonists & inhibitors
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CD40 Ligand / blood
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CD40 Ligand / metabolism*
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Case-Control Studies
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Cell-Derived Microparticles / enzymology*
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Cell-Derived Microparticles / pathology
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Cells, Cultured
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Disease Models, Animal
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Disseminated Intravascular Coagulation / enzymology
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Disseminated Intravascular Coagulation / pathology
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Endothelial Cells / drug effects
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Endothelial Cells / enzymology*
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Endothelial Cells / pathology
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Endotoxemia / enzymology
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Endotoxemia / genetics
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Endotoxemia / pathology
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Female
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Gene Expression Regulation, Enzymologic
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Hirudins / pharmacology
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Human Umbilical Vein Endothelial Cells / enzymology
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Humans
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Lipopolysaccharides / pharmacology
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Male
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Matrix Metalloproteinase 10 / blood
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Matrix Metalloproteinase 10 / deficiency
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Matrix Metalloproteinase 10 / genetics
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Matrix Metalloproteinase 10 / metabolism*
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Middle Aged
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Mitogen-Activated Protein Kinase 8 / antagonists & inhibitors
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Mitogen-Activated Protein Kinase 8 / metabolism
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Multivariate Analysis
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Peptides / pharmacology
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Protein Kinase Inhibitors / pharmacology
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RNA, Messenger / metabolism
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Receptor, PAR-1 / agonists
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Receptor, PAR-1 / antagonists & inhibitors
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Receptor, PAR-1 / metabolism
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Risk Assessment
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Risk Factors
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Sepsis / enzymology*
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Sepsis / mortality
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Sepsis / pathology
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Signal Transduction
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Spain
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Survival Analysis
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Thrombin / metabolism*
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Time Factors
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Up-Regulation
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p38 Mitogen-Activated Protein Kinases / antagonists & inhibitors
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p38 Mitogen-Activated Protein Kinases / metabolism
Substances
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Antibodies
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CD40 Antigens
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Hirudins
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Lipopolysaccharides
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Peptides
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Protein Kinase Inhibitors
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RNA, Messenger
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Receptor, PAR-1
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CD40 Ligand
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Mitogen-Activated Protein Kinase 8
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p38 Mitogen-Activated Protein Kinases
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Thrombin
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MMP10 protein, human
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Matrix Metalloproteinase 10