Dual regulatory effects of nitric oxide on plasminogen activator inhibitor type 1 expression in endothelial cells.
In this report we compared the mechanism by which nitric oxide (NO), generated exogenously and endogenously, affects the plasminogen activator inhibitor type 1 (PAI-1) expression in endothelial cells. For this purpose, we stimulated the endothelial cell line EA.hy 926 with tumour necrosis factor alpha (TNFalpha) in the presence of the exogenously NO-releasing donors, sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine, or regulators of nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine-methyl ester hydrochloride and substrate L-Arg. Expression of PAI-1 in EA.hy 926 cells was determined by measuring the level of mRNA, using relative quantitative reverse transcriptase PCR, and protein, using ELISA. In addition, we estimated the level of activation of two mitogen-activated protein kinases (MAPKs), extracellular signal-regulated kinase (ERK1/2) and c-Jun N-terminal kinase (JNK1/2), in the cells before and after treatment with TNFalpha, in the presence or absence of NO donors and inhibitors. In contrast to exogenously released NO that significantly reduced mostly basal PAI-1 expression, endogenously generated NO by NOS potentiated TNFalpha-induced upregulation of PAI-1 expression. Exogenously and endogenously generated NO causes different effects on activation of the MAPKs ERK1/2 and JNK1/2. Specifically, the SNP-released NO activates only ERK1/2, while endogenously generated NO in a pathway induced by TNFalpha activates both MAPKs. Thus our data indicate that due to different cellular locations and mechanisms of generation, NO may participate in various signalling pathways leading to opposite effects on PAI-1 expression in endothelial cells.
Swiatkowska
M
M
Department of Biophysics, Medical University in Lodz; Center for Microbiology and Virology, Polish Academy of Sciences, Lodz, Poland.
Cierniewska-Cieslak
A
A
Pawlowska
Z
Z
Cierniewski
C S
CS
D001120:Arginine / Q000378:metabolism; D002460:Cell Line; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D005419:Flavonoids / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism* / Q000494:pharmacology; D020030:Nitric Oxide Donors / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects
D005419:Flavonoids; D020030:Nitric Oxide Donors; D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D001120:Arginine; D019001:Nitric Oxide Synthase; D020928:Mitogen-Activated Protein Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D019331:NG-Nitroarginine Methyl Ester