Mechanism of tissue factor production by monocytes stimulated with neutrophil elastase

Blood Cells Mol Dis. 2015 Feb;54(2):206-9. doi: 10.1016/j.bcmd.2014.10.005. Epub 2014 Oct 30.

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.

Aims: 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.

Keywords: Disseminated intravascular coagulation; Monocyte; Neutrophil elastase; Protein kinase C; Tissue factor.

MeSH terms

  • Calcium Channel Blockers / pharmacology
  • Carbazoles / pharmacology
  • Enzyme Inhibitors / pharmacology
  • Estrenes / pharmacology
  • Flavonoids / pharmacology
  • Gallic Acid / analogs & derivatives
  • Gallic Acid / pharmacology
  • Gene Expression Regulation
  • Humans
  • Imidazoles / pharmacology
  • Indoles / pharmacology
  • Leukocyte Elastase / pharmacology*
  • Maleimides / pharmacology
  • Monocytes / cytology
  • Monocytes / drug effects*
  • Monocytes / metabolism
  • Primary Cell Culture
  • Protein Kinase C / genetics*
  • Protein Kinase C / metabolism
  • Protein Kinase Inhibitors / pharmacology
  • Pyridines / pharmacology
  • Pyrrolidinones / pharmacology
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Signal Transduction
  • Thromboplastin / genetics*
  • Thromboplastin / metabolism
  • Type C Phospholipases / genetics*
  • Type C Phospholipases / metabolism
  • p38 Mitogen-Activated Protein Kinases / genetics*
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • 2-(1-(3-dimethylaminopropyl)-5-methoxyindol-3-yl)-3-(1H-indol-3-yl)maleimide
  • Calcium Channel Blockers
  • Carbazoles
  • Enzyme Inhibitors
  • Estrenes
  • Flavonoids
  • Imidazoles
  • Indoles
  • Maleimides
  • Protein Kinase Inhibitors
  • Pyridines
  • Pyrrolidinones
  • RNA, Messenger
  • 1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione
  • Ro 31-8425
  • Go 6976
  • 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate
  • Gallic Acid
  • Thromboplastin
  • Protein Kinase C
  • p38 Mitogen-Activated Protein Kinases
  • Type C Phospholipases
  • Leukocyte Elastase
  • SB 203580
  • 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one