title: Blockade by polyunsaturated n-3 fatty acids of endotoxin-induced monocytic tissue factor activation is mediated by the depressed receptor expression in THP-1 cells.
abstract: BACKGROUND
  Monocytic hypercoagulation often occurs in inflammatory conditions. We have previously reported that polyunsaturated n-3 fatty acids (n-3 FA) including eicosapentaenoic acid (20:5) and docosahexaenoic acid (22:6) prevent the activation of monocytic tissue factor (TF) induced by bacterial endotoxin [lipopolysaccharide (LPS)] in cell cultures and animals.
  
  
  OBJECTIVE
  We herein explore the mode of inhibitory action of n-3 FA to determine if LPS transmembrane signaling is blocked, exerting such antagonism.
  
  
  RESULTS
  Exposure of human leukemia monocytic THP-1 cells to bacterial endotoxin (Escherichia coli 0111:B04, 1.5 microg/ml) for 6 h significantly activated TF activity and the production of nitric oxide (NO), tumor necrosis factor alpha (TNF-alpha), and interleukin (IL)-1beta in conditioned medium. Pretreatment with n-3 FA, 20:5 and 22:6 at 10 microM, resulted in time-dependent suppression of not only TF activation but also the elicitation of NO, TNF-alpha, and IL-1beta. These LPS responses were substantially depressed by more than 50% after a 72-h pretreatment. FACScan analysis showed that n-3 FA readily prevented fluorescein isothiocyanate (FITC)-conjugated LPS from binding to THP-1 cells by approximately 70%. The observation that anti-CD14 mAb diminished FITC-LPS binding in a dose-dependent fashion has revealed CD14 dependency in LPS recognition. LPS upregulated CD14 expression, which was significantly arrested by n-3 FA. Similarly, the upregulation of the expression of CD11b, another proposed LPS receptor, was also minimally but significantly depressed by n-3 FA.
  
  
  CONCLUSIONS
  The present study demonstrates that n-3 FA are able to block LPS transmembrane signaling via suppression of the receptor upregulation, mediating a variety of significant antagonisms against LPS action.
authors.affiliation: School of Medicine, Wayne State University, Detroit, Michigan 48201, USA. ad5742@wayne.edu
authors.forename: A J
authors.initials: AJ
authors.lastname: Chu
authors.forename: M A
authors.initials: MA
authors.lastname: Walton
authors.forename: J K
authors.initials: JK
authors.lastname: Prasad
authors.forename: A
authors.initials: A
authors.lastname: Seto
mesh_terms: D018845:CD11 Antigens / Q000032:analysis; D015525:Fatty Acids, Omega-3 / Q000494:pharmacology*; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000032:analysis / Q000187:drug effects*; D008070:Lipopolysaccharides / Q000633:toxicity*; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D013925:Thromboplastin / Q000096:biosynthesis*; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D018845:CD11 Antigens; D015525:Fatty Acids, Omega-3; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D013925:Thromboplastin