Download PDF
Thromb Haemost 2009; 101(06): 1060-1069
DOI: 10.1160/TH08-03-0164
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Inhibition by fibrates of plasminogen activator inhibitor type-1 expression in human adipocytes and preadipocytes

Andreas Zirlik
1   Department of Cardiology, University of Freiburg, Freiburg, Germany
,
Sandra Ernst
1   Department of Cardiology, University of Freiburg, Freiburg, Germany
,
Anne Leugers
1   Department of Cardiology, University of Freiburg, Freiburg, Germany
,
Florian Willecke
1   Department of Cardiology, University of Freiburg, Freiburg, Germany
,
Burton E. Sobel
2   Department of Medicine, University of Vermont College of Medicine, Burlington, Vermont, USA
,
Christoph Bode
1   Department of Cardiology, University of Freiburg, Freiburg, Germany
,
Thomas K. Nordt
3   Department of Cardiology, Katharinenhospital, Klinikum Stuttgart, Stuttgart, Germany
› Author AffiliationsFinancial support: This study was supported by grants from the University of Freiburg and the Deutsche Forschungsgemeinschaft to A.Z. (DFG ZI 743/1–1 and 3–1) and by a grant from Astra Zeneca to T.K.N.
Further Information

Publication History

Received: 14 March 2008

Accepted after major revision: 24 February 2009

Publication Date:
24 November 2017 (online)

    Permissions and Reprints

Summary

Plasminogen activator inhibitor type-1 (PAI-1), an established marker and mediator of cardiovascular risk, is produced extensively in adipose tissue. Fibrates are hypolipidemic peroxisome proliferator activated receptor-alpha (PPARα) agonists. Recent laboratory and clinical observations indicate that they are also anti-atherosclerotic. Mechanisms responsible, however, remain to be fully understood. The present study was designed to elucidate modulation of PAI-1 expression in adipose cells by fibrates as a potential mechanism. Expression of PPARα was verified by PCR, immunohistochemistry, and Western blotting. In cultured preadipocytes and adipocytes gemfibrozil and fenofibrate significantly reduced PAI-1 protein expression by up to 55 ± 5% and 34 ± 4% under basal conditions and up to 56 ± 6% and 31 ± 6% under conditions of stimulation of the cells with 40 pM trans-Keywords forming growth factor (TGF)β, respectively. Quantification of mRNA showed that the gemfibrozil-induced effect was at least in part regulated at the transcriptional level. Incubations with non-fibrate PPARα agonists showed similar reductions in PAI-1 expression. The decrease in PAI-1 expression induced by gemfibrozil was inhibited by MK886, a PPARα inhibitor. Furthermore, preadipocytes isolated from PPARα-deficient mice produced significantly more PAI-1 than those from wild-type mice upon stimulation with TGFβ. Finally, fenofibrate reduced PAI-1 expression both in plasma and adipose tissue of hyperlipidemic mice. Our data support the view that PPARα activation down-regulates PAI-expression in adipose cells that may contribute in part to the reduction in cardiovascular mortality seen with fibrates in clinical trials.

Keywords

PAI-1 - adipocyte - fibrate - metabolic syndrome - haemostasis
 

  • References

  • 1 Chapman MJ. Fibrates in 2003: therapeutic action in atherogenic dyslipidaemia and future perspectives.. Atherosclerosis 2003; 171: 1-13.
    CrossrefPubMedGoogle Scholar
  • 2 Willson TM. et al. The PPARs: from orphan receptors to drug discovery.. J Med Chem 2000; 43: 527-550.
    CrossrefPubMedGoogle Scholar
  • 3 Effect of fenofibrate on progression of coronary-artery disease in type 2 diabetes: the Diabetes Atherosclerosis Intervention Study, a randomised study.. Lancet 2001; 357: 905-910.
    CrossrefPubMedGoogle Scholar
  • 4 Ericsson CG. et al. Angiographic assessment of effects of bezafibrate on progression of coronary artery disease in young male postinfarction patients.. Lancet 1996; 347: 849-853.
    CrossrefPubMedGoogle Scholar
  • 5 Frick MH. et al. Helsinki Heart Study: primary-prevention trial with gemfibrozil in middle-aged men with dyslipidemia. Safety of treatment, changes in risk factors, and incidence of coronary heart disease.. N Engl J Med 1987; 317: 1237-1245.
    CrossrefPubMedGoogle Scholar
  • 6 Keech A. et al. Effects of long-term fenofibrate therapy on cardiovascular events in 9795 people with type 2 diabetes mellitus (the FIELD study): randomised controlled trial.. Lancet 2005; 366: 1849-1861.
    CrossrefPubMedGoogle Scholar
  • 7 Robins SJ. et al. Relation of gemfibrozil treatment and lipid levels with major coronary events: VA-HIT: a randomized controlled trial.. J Am Med Assoc 2001; 285: 1585-1591.
    CrossrefPubMedGoogle Scholar
  • 8 Rubins HB. et al. Gemfibrozil for the secondary prevention of coronary heart disease in men with low levels of high-density lipoprotein cholesterol. Veterans Affairs High-Density Lipoprotein Cholesterol Intervention Trial Study Group.. N Engl J Med 1999; 341: 410-418.
    CrossrefPubMedGoogle Scholar
  • 9 Bairaktari ET. et al. Comparison of the efficacy of atorvastatin and micronized fenofibrate in the treatment of mixed hyperlipidemia.. J Cardiovasc Risk 1999; 6: 113-116.
    CrossrefPubMedGoogle Scholar
  • 10 Marx N. et al. PPARalpha activators inhibit tissue factor expression and activity in human monocytes.. Circulation 2001; 103: 213-219.
    CrossrefPubMedGoogle Scholar
  • 11 Marx N. et al. PPARalpha activators inhibit cytokine-induced vascular cell adhesion molecule-1 expression in human endothelial cells.. Circulation 1999; 99: 3125-3131.
    CrossrefPubMedGoogle Scholar
  • 12 Wang TD. et al. Efficacy of fenofibrate and simvastatin on endothelial function and inflammatory markers in patients with combined hyperlipidemia: relations with baseline lipid profiles.. Atherosclerosis 2003; 170: 315-323.
    CrossrefPubMedGoogle Scholar
  • 13 Kohler HP, Grant PJ. Plasminogen-activator inhibitor type 1 and coronary artery disease.. N Engl J Med 2000; 342: 1792-1801.
    CrossrefPubMedGoogle Scholar
  • 14 Tsantes AE. et al. Association between the plasminogen activator inhibitor-1 4G/5G polymorphism and venous thrombosis. A meta-analysis.. Thromb Haemost 2007; 97: 907-913.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 15 Lyon CJ, Hsueh WA. Effect of plasminogen activator inhibitor-1 in diabetes mellitus and cardiovascular disease.. Am J Med 2003; 115 Suppl 8A 62S-68S.
    PubMedGoogle Scholar
  • 16 Sobel BE. The potential influence of insulin and plasminogen activator inhibitor type 1 on the formation of vulnerable atherosclerotic plaques associated with type 2 diabetes.. Proc Assoc Am Physicians 1999; 111: 313-318.
    CrossrefPubMedGoogle Scholar
  • 17 Kinik ST. et al. PAI-1 gene 4G/5G polymorphism, cytokine levels and their relations with metabolic parameters in obese children.. Thromb Haemost 2008; 99: 352-356.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 18 Lundgren CH. et al. Elaboration of type-1 plasminogen activator inhibitor from adipocytes. A potential pathogenetic link between obesity and cardiovascular disease.. Circulation 1996; 93: 106-110.
    CrossrefPubMedGoogle Scholar
  • 19 Morange PE. et al. PAI-1 produced ex vivo by human adipose tissue is relevant to PAI-1 blood level.. Arterioscler Thromb Vasc Biol 1999; 19: 1361-1365.
    CrossrefPubMedGoogle Scholar
  • 20 Bastelica D. et al. Stromal cells are the main plasminogen activator inhibitor-1-producing cells in human fat: evidence of differences between visceral and subcutaneous deposits.. Arterioscler Thromb Vasc Biol 2002; 22: 173-178.
    CrossrefPubMedGoogle Scholar
  • 21 Bastelica D. et al. Relationships between fibrinolytic and inflammatory parameters in human adipose tissue: strong contribution of TNFalpha receptors to PAI-1 levels.. Thromb Haemost 2002; 88: 481-487.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 Folsom AR. et al. Impact of weight loss on plasminogen activator inhibitor (PAI-1), factor VII, and other hemostatic factors in moderately overweight adults.. Arterioscler Thromb 1993; 13: 162-169.
    CrossrefPubMedGoogle Scholar
  • 23 Primrose JN. et al. Reduction in factor VII, fibrinogen and plasminogen activator inhibitor-1 activity after surgical treatment of morbid obesity.. Thromb Haemost 1992; 68: 396-399.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 24 Birgel M. et al. Role of cytokines in the regulation of plasminogen activator inhibitor-1 expression and secretion in newly differentiated subcutaneous human adipocytes.. Arterioscler Thromb Vasc Biol 2000; 20: 1682-1687.
    CrossrefPubMedGoogle Scholar
  • 25 Alessi MC, Juhan-Vague I. Metabolic syndrome, haemostasis and thrombosis.. Thromb Haemost 2008; 99: 995-1000.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 26 Zirlik A. et al. Direct attenuation of plasminogen activator inhibitor type-1 expression in human adipose tissue by thiazolidinediones.. Thromb Haemost 2004; 91: 674-682.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 27 Staels B. et al. Activation of human aortic smooth-muscle cells is inhibited by PPARalpha but not by PPARgamma activators.. Nature 1998; 393: 790-793.
    CrossrefPubMedGoogle Scholar
  • 28 Zirlik A. et al. TRAF-1, –2, –3, –5, and –6 are induced in atherosclerotic plaques and differentially mediate proinflammatory functions of CD40L in endothelial cells.. Arterioscler Thromb Vasc Biol 2007; 27: 1101-1107.
    CrossrefPubMedGoogle Scholar
  • 29 Zirlik A. et al. CD40 ligand mediates inflammation independently of CD40 by interaction with Mac-1.. Circulation 2007; 115: 1571-1580.
    CrossrefPubMedGoogle Scholar
  • 30 Gorzelniak K. et al. Validation of endogenous controls for gene expression studies in human adipocytes and preadipocytes.. Horm Metab Res 2001; 33: 625-627.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 31 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.. Methods 2001; 25: 402-408.
    CrossrefPubMedGoogle Scholar
  • 32 Bernlohr DA. et al. Evidence for an increase in transcription of specific mRNAs during differentiation of 3T3-L1 preadipocytes.. J Biol Chem 1985; 260: 5563-5567.
    PubMedGoogle Scholar
  • 33 Beranek JT. CD68 is not a macrophage-specific antigen.. Ann Rheum Dis 2005; 64: 342-343 author reply 343-344.
    PubMedGoogle Scholar
  • 34 Khazen W. et al. Expression of macrophage-selective markers in human and rodent adipocytes.. FEBS Lett 2005; 579: 5631-5634.
    CrossrefPubMedGoogle Scholar
  • 35 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
  • 36 Kruszynska YT. et al. Effects of troglitazone on blood concentrations of plasminogen activator inhibitor 1 in patients with type 2 diabetes and in lean and obese normal subjects.. Diabetes 2000; 49: 633-639.
    CrossrefPubMedGoogle Scholar
  • 37 Sobel BE. Increased plasminogen activator inhibitor-1 and vasculopathy. A reconcilable paradox.. Circulation 1999; 99: 2496-2498.
    CrossrefPubMedGoogle Scholar
  • 38 Vague P. et al. Correlation between blood fibrinolytic activity, plasminogen activator inhibitor level, plasma insulin level, and relative body weight in normal and obese subjects.. Metabolism Clin Exp 1986; 35: 250-253.
    CrossrefPubMedGoogle Scholar
  • 39 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
  • 40 Kockx M. et al. Studies on the role of PPAR in the fibrate-modulated gene expression of apolipoprotein A-I, plasminogen activator inhibitor 1, and fibrinogen in primary hepatocyte cultures from cynomolgus monkey.. Ann NY Acad Sci 1996; 804: 711-712.
    CrossrefPubMedGoogle Scholar
  • 41 Arts J. et al. Studies on the mechanism of fibrate-inhibited expression of plasminogen activator inhibitor-1 in cultured hepatocytes from cynomolgus monkey.. Arterioscler Thromb Vasc Biol 1997; 17: 26-32.
    CrossrefPubMedGoogle Scholar
  • 42 Nordt TK. et al. Attenuation by fibrates of plasminogen activator inhibitor type-1 expression in human arterial smooth muscle cells.. Thromb Haemost 2001; 86: 1305-1313.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 43 Schneider DJ, Sobel BE. Synergistic augmentation of expression of plasminogen activator inhibitor type-1 induced by insulin, very-low-density lipoproteins, and fatty acids.. Coron Artery Dis 1996; 7: 813-817.
    CrossrefPubMedGoogle Scholar
  • 44 Sobel BE. et al. Intramural plasminogen activator inhibitor type-1 and coronary atherosclerosis.. Arterioscler Thromb Vasc Biol 2003; 23: 1979-1989.
    CrossrefPubMedGoogle Scholar
  • 45 Zhao SP. et al. Fenofibrate inhibits thrombogenic and fibrinolytic factors expression in adipose tissue of atherosclerotic rabbits.. Clin Chim Acta 2004; 349: 81-86.
    CrossrefPubMedGoogle Scholar
  • 46 Avellone G. et al. Improvement of fibrinolysis and plasma lipoprotein levels induced by gemfibrozil in hypertriglyceridemia.. Blood Coagul Fibrinolysis 1995; 6: 543-548.
    CrossrefPubMedGoogle Scholar
  • 47 Okopien B. et al. Hypolipidemic drugs affect monocyte IL-1beta gene expression and release in patients with IIa and IIb dyslipidemia.. J Cardiovasc Pharmacol 2005; 45: 160-164.
    CrossrefPubMedGoogle Scholar
  • 48 Broijersen A. et al. Gemfibrozil treatment of combined hyperlipoproteinemia. No improvement of fibrinolysis despite marked reduction of plasma triglyceride levels.. Arterioscler Thromb Vasc Biol 1996; 16: 511-516.
    CrossrefPubMedGoogle Scholar
  • 49 Hamsten A. et al. Fibrinolytic proteins and progression of coronary artery disease in relation to gemfibrozil therapy.. Thromb Haemost 2000; 83: 397-403.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 50 Kockx M. et al. Effects of gemfibrozil and ciprofibrate on plasma levels of tissue-type plasminogen activator, plasminogen activator inhibitor-1 and fibrinogen in hyperlipidaemic patients.. Thromb Haemost 1997; 78: 1167-1172.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 51 Cimminiello C. et al. Fibrinolytic response in subjects with hypertriglyceridemia and low HDL cholesterol.. Biomed Pharmacother 1997; 51: 164-169.
    CrossrefPubMedGoogle Scholar
  • 52 Kehrer JP. et al. Inhibition of peroxisome-proliferator-activated receptor (PPAR)alpha by MK886.. Biochem J 2001; 356: 899-906.
    CrossrefPubMedGoogle Scholar
  • 53 Mavri A. et al. PPARalpha deficiency does not modify age dependency but prevents high fat diet increase in plasma PAI-1 as well as insulin resistance.. Thromb Haemost 2004; 91: 1051-1052.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 54 Guerre-Millo M. et al. PPAR-alpha-null mice are protected from high-fat diet-induced insulin resistance.. Diabetes 2001; 50: 2809-2814.
    CrossrefPubMedGoogle Scholar
  • 55 Patel DD. et al. Disturbances in the normal regulation of SREBP-sensitive genes in PPAR alpha-deficient mice.. J Lipid Res 2001; 42: 328-337.
    PubMedGoogle Scholar
  • 56 Delerive P. et al. Peroxisome proliferator-activated receptor activators inhibit thrombin-induced endothelin-1 production in human vascular endothelial cells by inhibiting the activator protein-1 signaling pathway.. Circulation Res 1999; 85: 394-402.
    CrossrefPubMedGoogle Scholar
  • 57 Calkin AC. et al. Gemfibrozil decreases atherosclerosis in experimental diabetes in association with a reduction in oxidative stress and inflammation.. Diabetologia 2006; 49: 766-774.
    CrossrefPubMedGoogle Scholar
  • 58 Hennuyer N. et al. PPARalpha, but not PPARgamma, activators decrease macrophage-laden athero-sclerotic lesions in a nondiabetic mouse model of mixed dyslipidemia.. Arterioscler Thromb Vasc Biol 2005; 25: 1897-1902.
    CrossrefPubMedGoogle Scholar
  • 59 Kooistra T. et al. Fenofibrate reduces atherogenesis in ApoE*3Leiden mice: evidence for multiple antiatherogenic effects besides lowering plasma cholesterol.. Arterioscler Thromb Vasc Biol 2006; 26: 2322-2330.
    CrossrefPubMedGoogle Scholar
  • 60 Srivastava RA. et al. Peroxisome proliferator-activated receptor-alpha selective ligand reduces adiposity, improves insulin sensitivity and inhibits atherosclerosis in LDL receptor-deficient mice.. Mol Cell Biochem 2006; 285: 35-50.
    CrossrefPubMedGoogle Scholar
  • 61 Tabernero A. et al. Activation of the peroxisome proliferator-activated receptor alpha protects against myocardial ischaemic injury and improves endothelial vasodilatation.. BMC Pharmacol 2002; 2: 10.
    CrossrefPubMedGoogle Scholar
  • 62 Damci T. T. et al. Fenofibrate treatment is associated with better glycemic control and lower serum leptin and insulin levels in type 2 diabetic patients with hyper-triglyceridemia.. Eur J Intern Med 2003; 14: 357-360.
    CrossrefPubMedGoogle Scholar