title: Regulation of tissue factor expression in smooth muscle cells with nitric oxide.
abstract: OBJECTIVE
  This study was undertaken to determine the effect of nitric oxide (NO) on tissue factor (TF) expression in vascular smooth muscle cells.
  
  
  METHODS
  Rat aortic smooth muscle cells (RASMCs) were exposed to NO delivered exogenously with the NO donor S-nitroso-N-acetylpenicillamine (SNAP) or produced endogenously after infection with an adenoviral vector carrying human inducible NO synthase (AdiNOS). Functional TF activity was assessed with chromogenic TF assay. TF antigen was determined with immunohistochemistry. Northern blot analysis was used to determine steady- state TF messenger RNA (mRNA). Electrophoretic mobility gel shift assay was performed to determine the nuclear binding activity of nuclear factor kappa-B (NFkappaB). NFkappaB activity was inhibited by either prior transduction of RASMCs with mutant IkappaB or treatment with pyrrolidine dithiocarbamate.
  
  
  RESULTS
  RASMCs exposed to SNAP or infected with AdiNOS exhibited increased functional TF activity and antigen. Regardless of the source of NO, a time-dependent and concentration-dependent increase in TF activity was observed. Steady-state TF mRNA levels were also increased by NO delivered via either method. NFkappaB nuclear binding activity was also increased by NO. Inhibition of NFkappaB activity by either pyrrolidine dithiocarbamate treatment or mutant IkappaB transduction abrogated NO-induced enhancement of TF mRNA and functional activity.
  
  
  CONCLUSIONS
  In RASMC, NO exposure results in upregulation of TF functional activity, antigen, and mRNA. This effect appears to be mediated by an NFkappaB-dependent pathway.
authors.affiliation: Department of Surgery, A-1011, University of Pittsburgh, 200 Lothrop Street, Pittsburgh, PA 15213, USA.
authors.forename: Melina R
authors.initials: MR
authors.lastname: Kibbe
authors.forename: Christopher
authors.initials: C
authors.lastname: Johnnides
authors.forename: Susan
authors.initials: S
authors.lastname: Gleixner
authors.forename: Imre
authors.initials: I
authors.lastname: Kovesdi
authors.forename: Alena
authors.initials: A
authors.lastname: Lizonova
authors.forename: Brian
authors.initials: B
authors.lastname: Zuckerbraun
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Billiar
authors.forename: Edith
authors.initials: E
authors.lastname: Tzeng
authors.forename: Satish C
authors.initials: SC
authors.lastname: Muluk
mesh_terms: D000256:Adenoviridae; D000818:Animals; D000941:Antigens / Q000378:metabolism; D001013:Aorta, Thoracic; D001681:Biological Assay; D015152:Blotting, Northern; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005822:Genetic Vectors; D020672:I-kappa B Proteins / Q000235:genetics / Q000494:pharmacology; D007150:Immunohistochemistry; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D009569:Nitric Oxide / Q000494:pharmacology / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000235:genetics / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D011485:Protein Binding; D011759:Pyrrolidines / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D026423:S-Nitroso-N-Acetylpenicillamine / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D013925:Thromboplastin / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014162:Transfection; D015854:Up-Regulation
chemical_list: D000941:Antigens; D020672:I-kappa B Proteins; D016328:NF-kappa B; D020030:Nitric Oxide Donors; D011759:Pyrrolidines; D012333:RNA, Messenger; D013859:Thiocarbamates; C020972:pyrrolidine dithiocarbamic acid; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine; D004247:DNA; D013925:Thromboplastin; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat