title: Involvement of MAPK activation in bacterial endotoxin-inducible tissue factor upregulation in human monocytic THP-1 cells.
abstract: BACKGROUND
  Monocytic tissue factor (mTF) hypercoagulation leading to thrombotic complications is commonly observed following sepsis.
  
  
  OBJECTIVE
  We herein study the intracellular mechanism of mTF upregulation in human model monocytic THP-1 cells in response to bacterial endotoxin (lipopolysaccharide, LPS; Escherichia coli O111:B04), determining if mitogen-activated protein kinase (MAPK) activation is involved in the signaling.
  
  
  METHODS
  We assessed mTF upregulation by its cell surface expression, protein synthesis, and functional activity based on flow cytometry, Western blotting analysis, and a single-stage clotting assay, respectively.
  
  
  RESULTS
  A 3-h challenge with LPS (100 ng/ml) drastically induced mTF functional activity, accompanied by elevated surface mTF expression and synthesis. The suppression by genistein (G) of LPS-inducible mTF upregulation implied the involvement of protein tyrosine kinase activation in mTF upregulation. LPS activated MAPK, which was significantly depressed by G, SB 203580 (SB), and PD 98058 (PD). Interestingly, inclusion of SB and PD also markedly diminished LPS-inducible mTF upregulation. The parallelism between MAPK and mTF activities revealed the involvement of MAPK activation in such mTF upregulation. Based on the ability of SB and PD to respectively block LPS-inducible tyrosine phosphorylation of p38 MAPK and Erk1/2, it was evident that tyrosine phosphorylation of MAPKs is required for mediating LPS-inducible mTF synthesis and upregulation. Contrasting with the established prevention of mTF upregulation by these inhibitors, failure to offset the already LPS-induced mTF activity seemed to be consistent with the view that LPS readily activated MAPK responsible for mTF synthesis.
  
  
  CONCLUSIONS
  Our data suggest that the tyrosine phosphorylation of MAPKs (p38 and Erk1/2) leading to their activation could be a prerequisite for LPS induction of mTF synthesis contributing to the upregulation of mTF-initiated extrinsic coagulation.
authors.affiliation: Department of Surgery, Wayne State University, Detroit, Michigan 48201, USA. ad5742@wayne.edu
authors.forename: A J
authors.initials: AJ
authors.lastname: Chu
authors.forename: Z G
authors.initials: ZG
authors.lastname: Wang
authors.forename: M A
authors.initials: MA
authors.lastname: Walton
authors.forename: A
authors.initials: A
authors.lastname: Seto
mesh_terms: D002460:Cell Line; D004731:Endotoxins / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007527:Isoenzymes / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009000:Monocytes / Q000378:metabolism*; D010766:Phosphorylation; D011725:Pyridines / Q000494:pharmacology; D013925:Thromboplastin / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D004731:Endotoxins; D004791:Enzyme Inhibitors; D007093:Imidazoles; D007527:Isoenzymes; D008070:Lipopolysaccharides; D011725:Pyridines; D014443:Tyrosine; D013925:Thromboplastin; D019833:Genistein; D020928:Mitogen-Activated Protein Kinases; C093642:SB 203580