Influence of Nicotine and Cotinine on the Expression of Plasminogen Activator Activity in Bovine Aortic Endothelial Cells

1989 ◽  
Vol 61 (01) ◽  
pp. 070-076 ◽  
Author(s):  
Be-Sheng Kuo ◽  
Maciej Dryjski ◽  
Thorir D Bjornsson
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Dependent Manner ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Fibrin Plate ◽  
Sds Page ◽  
Aortic Endothelial

SummaryThe effects of nicotine and its major metabolite, cotinine, were evaluated on the secretion of plasminogen activator (PA) and plasminogen activator inhibitor (PAI) in cultured bovine aortic endothelial cells. Both compounds increased PA secretion, determined by 125I-fibrin plate assay, in a time- and dose-dependent manner. Maximum effects after 24 hr incubation were observed for nicotine at 10-8 M and for cotinine at 10-7 M, which corresponded to about 2.6-fold increases over control for both compounds. The pharmacological PA stimulation required both RNA and protein syntheses, as evidenced by inhibition by acfinomycin D and cycloheximide. Both control and treated cells produced multiple forms of PA, as evaluated by SDS-PAGE zymography, and a single form of PAI, as evidenced by reverse fibrin autography. Although activities of all species of PA were enhanced by nicotine and cotinine, these compounds had no significant effects on the release of PAI. These results thus suggest that nicotine and cotinine may have fibrinolytic activity in vivo.

ROLE OF POLYAMINES IN THE REGULATION OF SYNTHESIS AND SECRETION OF PLASMINOGEN ACTIVATOR FROM BOVINE AORTIC ENDOTHELIAL CELLS

1987 ◽  
Author(s):  
Be-Sheng Kuo ◽  
Gil Korner ◽  
Thorir D Bjornsson
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Polyamine Metabolism ◽  
Individual Species ◽  
Dependent Manner ◽  
Plate Method ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Fibrin Plate ◽  
Aortic Endothelial

The effects of three polyamines, putrescine (PUT), spermidine (SPD) and spermine (SPM), were investigated on the synthesis and secretion of plasminogen activator (PA) and antiactivator (PAI) activities in confluent bovine aortic endothelial cells. PA activity was determined bythe fibrin plate method, and individual species with PA and PAI activities were separated and visualized using SDS-PAGE with zymography and reverse fibrin autography. Both control cells and cells treated with polyamines secreted PA activity in a time-dependent fashion. After 24-hour incubation, the three polyamines enhanced PA secretion in a dose-dependent manner (10-6 to 2.5 × 10-3 M), with a potency order of SPM > SPD> PUT, as estimated by the fibrin plate method. The maximum PA releases after PUT (0.5 mM), SPD (2.5 mM) and SPM (0.5 mM) were 1.7, 4.5 and 5.4 times control levels, respectively. Concentrations lower than 1 μM had essentially no effects. The enhancement of PA activity by polyamines was blocked by actino-mycin D and cycloheximide, while it was not affected by inhibitors of polyamine biosynthesis except that the enhancement by PUT (0.5 mM) was reduced by methylglyoxal bis(guanylhydrazone). These data suggest that polyamines directly stimulate PA synthesis and secretion through promotion of gene transcription and translation, and that this effect appears to be related to their position in the biosynthetic pathway of polyamines. The kinetic patterns of activities of ornithine decarboxylase and S-adenosy-methionine decarboxylase in confluent endothelial cells stimulated withfresh culture medium suggest that there is rapid turnover of intracellular polyamines. Multiple forms of secreted PA were observed and both tissue- and urokinase-type PA were enhanced by polyamines, while the PAI activity, as evaluted by reverse fibrin autograpy, was apparently reduced. These experimental results suggest that polyamines may play an important role in the regulation of the synthesis and secretion of plasminogen activators, and that this biological function could be modified by disease states and by agents that are associated with altered polyamine metabolism.

EFFECTS OF NICOTINE AND COTININE ON THE SYNTHESIS AND RELEASE OF PLASMINOGEN ACTIVATOR FROM BOVINE AORTIC ENDOTHELIAL CELLS

1987 ◽  
Author(s):  
Be-Sheng Kuo ◽  
Maciej Dryjski ◽  
Thorir D Bjornsson
Keyword(s):  
Endothelial Cells ◽  
Cigarette Smoking ◽  
Plasminogen Activator ◽  
Fibrinolytic Activity ◽  
Dependent Manner ◽  
Individual Molecular Species ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Stimulation Of ◽  
Aortic Endothelial

While cigarette smoking has beenimplicated in the development of cardiovascular diseases, it has been reported to increase fibrinolytic activity in vivo. However, no data is available regarding the underlying mechanism of action. The present investigation was carried out to evaluate the effects of nicotine and its major metabolite, cotinine, on the seretion of plasminogen activator (PA) and PA inhibitor (PAI) by cultured bovine aortic endothelial cells. PA activity was determined by the fibrin plate method, and individual molecular species with PA and PAI activities were separated and visualized using SDS-PAGE with zymography and reverse fibrin autography. Both nicotine and cotinine increased PA secretion in a time- and dose-dependent manner. A maximum stimulation in PA secretion after 24-hour incubation was observed for nicotine at 10-8 M and for cotinine at 10-7> M, which corresponded to 2.5- and 2.7-foldincreases over control, respectively. These concentrations are in the range observed after cigarette smoking. The pharmacological stimulation of PA activity required both RNA and protein synthesis, as evidencedby its inhibition by cycloheximide and actinomycin D. Both control cells and cells treated with nicotine and cotinine produced multiple forms of PA and a single form of PAI. The PAI was mainly of the latent form as no quenching effect was observed on standard tissue plasminogen activator (tPA) and urokinase (UK) after they were mixed with the conditioned culture medium. The PAs werefound to consist of both tPA and UK,and the corresponding complexes with PAI, however, the UK bands were wider than the tPA bands. Both species were enhanced by nicotine and cotinine. Although activities of all species of PA were enhanced by nicotine and cotinine, these compounds had no apparent quantitative or qualitative effects on the release ofPAI. These results thus suggest that the mechanism underlying the enhanced fibrinolytic activity after cigarette smoking may be due to nicotine- and cotinine-induced stimulation of PA synthesis and subsequent release.

scholarly journals Extracellular matrix of cultured bovine aortic endothelial cells contains functionally active type 1 plasminogen activator inhibitor

Blood ◽  
1987 ◽  
Vol 70 (3) ◽  
pp. 721-728 ◽  
Author(s):  
J Mimuro ◽  
RR Schleef ◽  
DJ Loskutoff
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Activator Inhibitor ◽  
Pai 1 ◽  
Aortic Endothelial

The extracellular matrix (ECM) of cultured bovine aortic endothelial cells (BAEs) was analyzed by immunoblotting and reverse fibrin autography and shown to contain type 1 plasminogen activator inhibitor (PAI-1). Most PAI-1 in the ECM formed complexes with exogenously added tissue-type plasminogen activator (tPA), demonstrating that this PAI-1 was functionally active. The resulting tPA/PAI-1 complexes were recovered in the reaction solution, indicating that the PAI-1 in such complexes no longer bound to ECM. The PAI-1 could not be removed by incubating ECM in high salt (2 mol/L NaCl), sugars (1 mol/L galactose, 1 mol/L mannose), glycosaminoglycans (10 mmol/L heparin, 10 mmol/L dermatan sulfate), or epsilon-aminocaproic acid (0.1 mol/L). However, PAI-1 could be extracted from ECM by treatment with either arginine (0.5 mol/L) or potassium thiocyanate (2 mol/L), or by incubation under acidic conditions (pH 2.5). ECM depleted of PAI-1 by acid extraction was able to bind both the active and latent forms of PAI-1. In this instance, most of the bound PAI-1 did not form complexes with tPA, indicating that the latent form was not activated as a consequence of binding to ECM. Although the PAI-1 activity in conditioned medium decayed with a half-life (t 1/2) of less than 3 hours, the t 1/2 of ECM- associated PAI-1 was greater than 24 hours. These data suggest that PAI- 1 is produced by cultured BAEs in an active form and is then either released into the medium where it is rapidly inactivated or into the subendothelium where it binds to ECM. The specific binding of PAI-1 to ECM protects it from this inactivation.

scholarly journals Extracellular matrix of cultured bovine aortic endothelial cells contains functionally active type 1 plasminogen activator inhibitor

Blood ◽  
1987 ◽  
Vol 70 (3) ◽  
pp. 721-728 ◽  
Author(s):  
J Mimuro ◽  
RR Schleef ◽  
DJ Loskutoff
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Activator Inhibitor ◽  
Pai 1 ◽  
Aortic Endothelial

Abstract The extracellular matrix (ECM) of cultured bovine aortic endothelial cells (BAEs) was analyzed by immunoblotting and reverse fibrin autography and shown to contain type 1 plasminogen activator inhibitor (PAI-1). Most PAI-1 in the ECM formed complexes with exogenously added tissue-type plasminogen activator (tPA), demonstrating that this PAI-1 was functionally active. The resulting tPA/PAI-1 complexes were recovered in the reaction solution, indicating that the PAI-1 in such complexes no longer bound to ECM. The PAI-1 could not be removed by incubating ECM in high salt (2 mol/L NaCl), sugars (1 mol/L galactose, 1 mol/L mannose), glycosaminoglycans (10 mmol/L heparin, 10 mmol/L dermatan sulfate), or epsilon-aminocaproic acid (0.1 mol/L). However, PAI-1 could be extracted from ECM by treatment with either arginine (0.5 mol/L) or potassium thiocyanate (2 mol/L), or by incubation under acidic conditions (pH 2.5). ECM depleted of PAI-1 by acid extraction was able to bind both the active and latent forms of PAI-1. In this instance, most of the bound PAI-1 did not form complexes with tPA, indicating that the latent form was not activated as a consequence of binding to ECM. Although the PAI-1 activity in conditioned medium decayed with a half-life (t 1/2) of less than 3 hours, the t 1/2 of ECM- associated PAI-1 was greater than 24 hours. These data suggest that PAI- 1 is produced by cultured BAEs in an active form and is then either released into the medium where it is rapidly inactivated or into the subendothelium where it binds to ECM. The specific binding of PAI-1 to ECM protects it from this inactivation.

scholarly journals Urea transporters are distributed in endothelial cells and mediate inhibition of l-arginine transport

2002 ◽  
Vol 283 (3) ◽  
pp. F578-F582 ◽  
Author(s):  
Laszlo Wagner ◽  
Janet D. Klein ◽  
Jeff M. Sands ◽  
Chris Baylis
Keyword(s):  
Endothelial Cells ◽  
Cell Types ◽  
Inhibitory Effect ◽  
Rat Aorta ◽  
Wide Distribution ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Elevated Blood Urea Nitrogen ◽  
Aortic Endothelial

Our laboratory previously reported that uremic levels of urea inhibitl-arginine (l-Arg) transport into endothelial cells. The present study further investigated this effect. We measuredl-Arg transport in cultured bovine aortic endothelial cells with normal or high urea (25 mM). The urea transport inhibitor phloretin abolished the inhibitory effect of urea on l-Arg transport, suggesting a role for urea transporters (UTs). We screened bovine aortic endothelial cells and several other endothelial cell types for the presence of UTs by using Western blot analysis. UT-B was present in all endothelial cells, irrespective of species or location of derivation, whereas UT-A distribution was variable and sparse. UT-B was also abundant in rat aorta, mesenteric blood vessels, and spinotrapezius muscle, whereas UT-A distribution was, again, variable and sparse. Chronic elevation of urea had variable, inconsistent effects on UT abundance. This study showed that urea must enter endothelial cells, probably by UT-B, to inhibit l-Arg transport. In view of the wide distribution of UT-B in rat vasculature, elevated blood urea nitrogen may lead to endothelial l-Arg deficiency in vivo.

Sign in / Sign up

Export Citation Format

Share Document