Regulation of tissue factor expression in smooth muscle cells with nitric oxide.
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.
Kibbe
Melina R
MR
Department of Surgery, A-1011, University of Pittsburgh, 200 Lothrop Street, Pittsburgh, PA 15213, USA.
Johnnides
Christopher
C
Gleixner
Susan
S
Kovesdi
Imre
I
Lizonova
Alena
A
Zuckerbraun
Brian
B
Billiar
Timothy R
TR
Tzeng
Edith
E
Muluk
Satish C
SC
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
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