title: Synergistic effect of thrombin and CD40 ligand on endothelial matrix metalloproteinase-10 expression and microparticle generation in vitro and in vivo.
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.
  
  
  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.
authors.affiliation: Laboratory of Atherothrombosis, Division of Cardiovascular Sciences, CIMA, Av. Pio XII, 55, 31008 Pamplona, Navarra, Spain.
authors.forename: Sara
authors.initials: S
authors.lastname: Martínez de Lizarrondo
authors.forename: Carmen
authors.initials: C
authors.lastname: Roncal
authors.forename: Olivier
authors.initials: O
authors.lastname: Calvayrac
authors.forename: Cristina
authors.initials: C
authors.lastname: Rodríguez
authors.forename: Nerea
authors.initials: N
authors.lastname: Varo
authors.forename: Ana
authors.initials: A
authors.lastname: Purroy
authors.forename: Leonardo
authors.initials: L
authors.lastname: Lorente
authors.forename: José A
authors.initials: JA
authors.lastname: Rodríguez
authors.forename: Loïc
authors.initials: L
authors.lastname: Doeuvre
authors.forename: Sandra
authors.initials: S
authors.lastname: Hervás-Stubbs
authors.forename: Eduardo
authors.initials: E
authors.lastname: Angles-Cano
authors.forename: José A
authors.initials: JA
authors.lastname: Páramo
authors.forename: José
authors.initials: J
authors.lastname: Martínez-González
authors.forename: Josune
authors.initials: J
authors.lastname: Orbe
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001777:Blood Coagulation; D019013:CD40 Antigens / Q000037:antagonists & inhibitors / Q000378:metabolism; D023201:CD40 Ligand / Q000037:antagonists & inhibitors / Q000097:blood / Q000378:metabolism*; D016022:Case-Control Studies; D055252:Cell-Derived Microparticles / Q000201:enzymology* / Q000473:pathology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004211:Disseminated Intravascular Coagulation / Q000201:enzymology / Q000473:pathology; D042783:Endothelial Cells / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D019446:Endotoxemia / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006629:Hirudins / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000201:enzymology; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D053506:Matrix Metalloproteinase 10 / Q000097:blood / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008875:Middle Aged; D048055:Mitogen-Activated Protein Kinase 8 / Q000037:antagonists & inhibitors / Q000378:metabolism; D015999:Multivariate Analysis; D010455:Peptides / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D044463:Receptor, PAR-1 / Q000819:agonists / Q000378:metabolism; D018570:Risk Assessment; D012307:Risk Factors; D018805:Sepsis / Q000201:enzymology* / Q000401:mortality / Q000473:pathology; D015398:Signal Transduction; D013030:Spain; D016019:Survival Analysis; D013917:Thrombin / Q000378:metabolism*; D013997:Time Factors; D015854:Up-Regulation; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D010975:Platelet Aggregation Inhibitors
chemical_list: D000906:Antibodies; D019013:CD40 Antigens; D006629:Hirudins; D008070:Lipopolysaccharides; D010455:Peptides; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D044463:Receptor, PAR-1; D023201:CD40 Ligand; D048055:Mitogen-Activated Protein Kinase 8; D048051:p38 Mitogen-Activated Protein Kinases; D013917:Thrombin; C511770:MMP10 protein, human; D053506:Matrix Metalloproteinase 10; D010975:Platelet Aggregation Inhibitors