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Thromb Haemost 1993; 70(04): 642-647
DOI: 10.1055/s-0038-1649642
Original Article
Fibrinolysis
Schattauer GmbH Stuttgart

Inhibition of Endothelial Cell Expression of Plasminogen Activator Inhibitor Type-1 by Gemfibrozil

Satoshi Fujii
The Washington University School of Medicine, St. Louis, Missouri, USA
,
Hirofumi Sawa
The Washington University School of Medicine, St. Louis, Missouri, USA
,
Burton E Sobel
The Washington University School of Medicine, St. Louis, Missouri, USA
› Author Affiliations
Further Information

Publication History

Received 08 January 1993

Accepted after revision 01 June 1993

Publication Date:
05 July 2018 (online)

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Summary

Increased concentrations of plasminogen activator inhibitor type-1 (PAI-1) in plasma are associated with impaired fibrinolysis and venous and arterial thrombo-embolic disease. In pilot studies designed to identify pharmacologic approaches capable of diminishing such increases, we found that gemfibrozil attenuated the Stimulation of synthesis of PAI-1 in a human, immortal, hepatoma cell line (Hep G2) induced by platelets. The present study was performed to determine whether it exerts analogous effects in non-immortal endothelial cells and whether it may therefore facilitate fibrinolysis locally in vivo. Human umbilical vein endothelial cells were incubated with pharmacologic concentrations of gemfibrozil. Gemfibrozil, 100 μM, suppressed basal PAI-1 production by 15% and attenuated the augmentation of synthesis of PAI-1 induced by lysates from platelets (4 × 107/ml) by 36% over 24 h without inhibiting overall protein synthesis. In addition, the increases in PAI-1 mRNA otherwise induced by platelet lysates over 6 h were suppressed by 49% (Northern blots) without any demonstrable change in the intracellular half-life of PAI-1 mRNA. Pulse-chase experiments demonstrated diminution of PAI-1 protein synthesis in parallel with the changes observed in PAI-1 mRNA. To determine whether these effects of gemfibrozil on endothelial cells in vitro were paralleled by consistent changes in the concentrations of PAI-1 in plasma in vivo, we studied rabbits with induced carotid artery thrombosis and thrombolysis. The increases in plasma PAI-1 induced by thrombosis and thrombolysis were inhibited by gemfibrozil (8.75 mg/kg i.v.), by 14% and 75%, respectively, Increases in endothelial cell PAI-1 gene expression in aorta, liver, and heart (4–6 fold over control) induced by thrombolysis were attenuated by gemfibrozil as well. Thus, gemfibrozil can attenuate shut down of the fibrinolytic system that is known to accompany thrombosis and thrombolysis and may therefore diminish the likelihood of early, thrombotic coronary artery reocclusion following revascularization with fibrinolytic agents.

 

  • References

  • 1 Loskutoff DJ, Sawdey M, Mimuro J. Type 1 plasminogen activator inhibitor. In: Progress in Hemostasis and Thrombosis. Coller B. (ed) Philadelphia, PA: WB Saunders; 1989. 9 87-115
  • 2 Hamsten A, Wiman B, De Faire U, Blombäck M. Increased plasma levels of a rapid inhibitor of tissue plasminogen activator in young survivors of myocardial infarction. N Engl J Med 1985; 313: 1557-1563
    CrossrefPubMedGoogle Scholar
  • 3 Heikki FrickM, Elo O, Haapa K, Heinonen OP, Heinsalmi P, Helo P, Huttunen JK, Kaitaniemi P, Koskinen P, Manninen V, Mäenpää H, Mälkönen M, Mänttäri M, Norola S, Pasternack A, Pikkarainen J, Romo M, Sjöblom T, Nikkilä E. Helsinki Heart Study: Primary-prevention trial with gemfibrozil in middle-aged men with dyslipidemia. N Engl J Med 1987; 317: 1237-1245
    CrossrefPubMedGoogle Scholar
  • 4 Manninen V, Elo O, Heikki FrickM, Haapa K, Heinonen OP, Heinsalmi P, Helo P, Huttunen JK, Kaitaniemi P, Koskinen P, Mäenpää H, Mälkönen M, Mänttäri M, Norola S, Pasternack A, Pikkarainen J, Romo M, Sjöblom T, Nikkilä EA. Lipid alterations and decline in the incidence of coronary heart disease in the Helsinki Heart Study. JAMA 1988; 260: 641-651
    CrossrefPubMedGoogle Scholar
  • 5 Manninen V, Huttunen JK, Heinonen OP, Tenkanen L, Heikki FrickM. Relation between baseline lipid and lipoprotein values and the incidence of coronary heart disease in the Helsinki Heart Study. Am J Cardiol 1989; 63: 42H-47H
    CrossrefPubMedGoogle Scholar
  • 6 Frick MH, Manninen V, Huttunen JK, Heinonen OP, Tenkanen L, Mänttäri M. HDL-cholesterol as a risk factor in coronary heart disease: An update of the Helsinki Heart Study. Drugs 1990; (Suppl. 01) 40: 7-12
    CrossrefPubMedGoogle Scholar
  • 7 Anderson P, Smith P, Seljeflot I, Brataker S, Arnesen H. Effects of gemfibrozil on lipids and haemostasis after myocardial infarction. Thromb Haemostas 1990; 63: 174-177
    PubMedGoogle Scholar
  • 8 Lucore CL, Sobel BE. Interactions of tissue-type plasminogen activator with plasma inhibitors and their pharmacologic implications. Circulation 1988; 77: 660-669
    CrossrefPubMedGoogle Scholar
  • 9 Fujii S, Abendschein DR, Sobel BE. Augmentation of plasminogen activator inhibitor type 1 activity in plasma by thrombosis and by thrombolysis. J Am Coll Cardiol 1991; 18: 1547-1554
    CrossrefPubMedGoogle Scholar
  • 10 Schneider DJ, Nordt TK, Sobel BE. Stimulation by proinsulin of expression of plasminogen activator inhibitor type-1 in endothelial cells. Diabetes 1992; 41: 890-895
    CrossrefPubMedGoogle Scholar
  • 11 Fujii S, Sobel BE. Direct effects of gemfibrozil on the fibrinolytic system. Diminution of synthesis of plasminogen activator inhibitor type 1. Circulation 1992; 85: 1888-1893
    CrossrefPubMedGoogle Scholar
  • 12 Jaffe EA, Nachman RL, Becker CK, Minick CR. Culture of human endothelial cells derived from umbilical veins. J Clin Invest 1973; 52: 2745-2756
    CrossrefPubMedGoogle Scholar
  • 13 Fujii S, Hopkins WE, Sobel BE. Mechanisms contributing to increased synthesis of plasminogen activator inhibitor type-1 in endothelial cells by constituents of platelets and their implications for thrombolysis. Circulation 1991; 83: 645-651
    CrossrefPubMedGoogle Scholar
  • 14 Fujii S, Lucore CL, Hopkins WE, Billadello JJ, Sobel BE. Potential attenuation of fibrinolysis by growth factors released from platelets and their pharmacologic implications. Am J Cardiol 1989; 63: 1505-1511
    CrossrefPubMedGoogle Scholar
  • 15 Koli K, Lohi J, Hautanen A, Keski-Oja J. Enhancement of vitronectin expression in human HepG2 hepatoma cells by transforming growth factor-β1. Eur J Biochem 1991; 199: 337-345
    CrossrefPubMedGoogle Scholar
  • 16 Fujii S, Sobel BE. Induction of plasminogen activator inhibitor by products released from platelets. Circulation 1990; 82: 1485-1493
    CrossrefPubMedGoogle Scholar
  • 17 Fujii S, Sawa H, Saffitz JE, Lucore CL, Sobel BE. Induction of endothelial cell expression of the plasminogen activator inhibitor type 1 gene by thrombosis in vivo. Circulation 1992; 86: 2000-2010
    CrossrefPubMedGoogle Scholar
  • 18 Lucore CL, Fujii S, Wun T-C, Sobel BE, Billadello JJ. Regulation of the expression of type 1 plasminogen activator inhibitor in Hep G2 cells by epidermal growth factor. J Biol Chem 1988; 263: 15845-15848
    PubMedGoogle Scholar
  • 19 Thomas PS. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 1980; 77: 5201-5205
    CrossrefPubMedGoogle Scholar
  • 20 Colucci M, Paramo JA, Collen D. Generation in plasma of a fast-acting inhibitor of plasminogen activator in response to endotoxin Stimulation. J Clin Invest 1985; 75: 818-824
    CrossrefPubMedGoogle Scholar
  • 21 Masuda T, Yasue H, Ogawa H, Misumi I, Sakamoto T, Okubo H, Miyao Y, Kato H. Plasma plasminogen activator inhibitor activity and tissue plasminogen activator levels in patients with unstable angina and those with coronary spastic angina. Am Heart J 1992; 124: 314-319
    CrossrefPubMedGoogle Scholar
  • 22 Hopkins WE, Westerhausen Jr DR, Sobel BE, Billadello JJ. Transcriptional regulation of plasminogen activator inhibitor type-1 mRNA in Hep G2 cells by epidermal growth factor. Nucl Acids Res 1991; 19: 163-168
    CrossrefPubMedGoogle Scholar
  • 23 Westerhausen Jr DR, Hopkins WE, Billadello JJ. Multiple transforming growth factor-β-inducible elements regulate expression of the plasminogen activator inhibitor type-1 gene in Hep G2 cells. J Biol Chem 1991; 266: 1092-1100
    PubMedGoogle Scholar
  • 24 Shaw G, Kamen R. A conserved AU sequence from the 3‘ untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell 1986; 46: 659-667
    CrossrefPubMedGoogle Scholar
  • 25 Malter JS. Identification of an AUUUA-specific messenger RNA binding protein. Science 1989; 246: 664-666
    CrossrefPubMedGoogle Scholar
  • 26 Caput D, Beutler B, Hartog K, Thayer R, Brown-Shimer S, Cerami A. Identification of a common nucleotide sequence in the 3’- untranslated region of mRNA molecules specifying inflammatory mediators. Proc Natl Acad Sci USA 1986; 83: 1670-1674
    CrossrefPubMedGoogle Scholar
  • 27 Kruys V, Marinx O, Shaw G, Deschamps J, Huez G. Translational blockade imposed by cytokine-derived UA-rich sequences. Science 1989; 245: 852-855
    CrossrefPubMedGoogle Scholar
  • 28 Todd PA, Ward A. Gemfibrozil. A review of its pharmacodynamic and pharmacokinetic properties, and therapeutic use in dyslipidaemia. Drugs 1988; 36: 314-339
    CrossrefPubMedGoogle Scholar
  • 29 Preissner KT, Jenne D. Vitronectin: A new molecular connection in haemostasis. Thrombo Haemostas 1991; 66: 189-194
    PubMedGoogle Scholar
  • 30 Seiffert D, Keeton M, Eguchi Y, Sawdey M, Loskutoff DJ. Detection of vitronectin mRNA in tissues and cells of the mouse. Proc Natl Acad Sci USA 1991; 88: 9402-9406
    CrossrefPubMedGoogle Scholar
  • 31 Issemann I, Green S. Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators. Nature 1990; 347: 645-650
    CrossrefPubMedGoogle Scholar
  • 32 Reddy JK, Goel SK, Nemali MR, Carrino JJ, Laffler TG, Reddy MK, Sperbeck SJ, Osumi T, Hashimoto T, Lalwani ND, Rao MS. Transcriptional regulation of peroxisomal fatty acyl-CoA oxidase and enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase in rat liver by peroxisome proliferators. Proc Natl Acad Sci USA 1986; 83: 1747-1751
    CrossrefPubMedGoogle Scholar
  • 33 Tolbert NE. Metabolic pathways in peroxisomes and glyoxysomes. Annu Rev Biochem 1981; 50: 133-157
    CrossrefPubMedGoogle Scholar
  • 34 Nemali MR, Usuda N, Reddy MK, Oyasu K, Hashimoto T, Osumi T, Rao MS, Reddy JK. Comparison of constitutive and inducible levels of expression of peroxisomal β-oxidation and catalase genes in liver and extrahepatic tissues of rat. Can Res 1988; 48: 5316-5324
    PubMedGoogle Scholar
  • 35 Avellone G, DiGarbo V, Cordova R, Ranelli G, De Simone R, Bompiani G. Effect of gemfibrozil treatment on fibrinolysis system in patients with hypertriglyceridemia. Curr Therap Res 1992; 52: 338-348
    CrossrefPubMedGoogle Scholar