title: Mechanism of tissue factor production by monocytes stimulated with neutrophil elastase.
abstract: BACKGROUND
  Monocytes and neutrophils are activated during disseminated intravascular coagulation. Tissue factor, the main initiator of coagulation, is expressed by monocytes, while elastase is released by neutrophils.
  
  
  OBJECTIVE
  This study investigated tissue factor production by peripheral monocytes after stimulation with human neutrophil elastase.
  
  
  METHODS
  Tissue factor mRNA levels were investigated by the reverse transcriptase-polymerase chain reaction and tissue factor protein production was assessed by western blotting when monocytes were exposed to neutrophil elastase with or without preincubation using various inhibitors.
  
  
  RESULTS
  Neutrophil elastase upregulated tissue factor mRNA and protein levels in monocytes. Both U73122 (phospholipase C inhibitor) and TMB-8 (intracellular calcium antagonist) prevented the upregulation of tissue factor mRNA. SB203580 (p38 mitogen-activated protein kinase inhibitor) suppressed this response, but PD98059 (extracellular signal-regulated kinase inhibitor) did not. Ro-318425 (ATP-competitive and selective protein kinase C (PKC) inhibitor) and Go 6976 (inhibitor of conventional PKCs and PKCμ) blocked the upregulation of tissue factor mRNA expression. Go 6983 (broad-spectrum PKC inhibitor) and CGP 4125 (staurosporine analog) partially attenuated it, as did a PKC theta/delta inhibitor.
  
  
  CONCLUSIONS
  Neutrophil elastase mainly enhances tissue factor production by monocytes via the phospholipase C/conventional PKC/p38 MAPK pathway, although a novel PKC is also involved.
authors.affiliation: Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Jin
authors.initials: J
authors.lastname: Kawata
authors.affiliation: Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Aoki
authors.affiliation: Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Yasuji
authors.initials: Y
authors.lastname: Ishimaru
authors.affiliation: Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Tomomichi
authors.initials: T
authors.lastname: Ono
authors.affiliation: Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Katsurou
authors.initials: K
authors.lastname: Sagara
authors.affiliation: Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Narahara
authors.affiliation: Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Tamami
authors.initials: T
authors.lastname: Matsmoto
authors.affiliation: Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Hirose
authors.affiliation: Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan. Electronic address: yamaguti@kumamoto-hsu.ac.jp.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Yamaguchi
mesh_terms: D002121:Calcium Channel Blockers / Q000494:pharmacology; D002227:Carbazoles / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004963:Estrenes / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D005707:Gallic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D005786:Gene Expression Regulation; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D019272:Leukocyte Elastase / Q000494:pharmacology*; D008301:Maleimides / Q000494:pharmacology; D009000:Monocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D061251:Primary Cell Culture; D011493:Protein Kinase C / Q000235:genetics* / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013925:Thromboplastin / Q000235:genetics* / Q000378:metabolism; D010738:Type C Phospholipases / Q000235:genetics* / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism
chemical_list: C465664:2-(1-(3-dimethylaminopropyl)-5-methoxyindol-3-yl)-3-(1H-indol-3-yl)maleimide; D002121:Calcium Channel Blockers; D002227:Carbazoles; D004791:Enzyme Inhibitors; D004963:Estrenes; D005419:Flavonoids; D007093:Imidazoles; D007211:Indoles; D008301:Maleimides; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D011760:Pyrrolidinones; D012333:RNA, Messenger; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; C071924:Ro 31-8425; C081021:Go 6976; C006014:8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate; D005707:Gallic Acid; D013925:Thromboplastin; D011493:Protein Kinase C; D048051:p38 Mitogen-Activated Protein Kinases; D010738:Type C Phospholipases; D019272:Leukocyte Elastase; C093642:SB 203580; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
keywords: Disseminated intravascular coagulation; Monocyte; Neutrophil elastase; Protein kinase C; Tissue factor