Platelet tissue factor activity and membrane cholesterol are increased in hypercholesterolemia and normalized by rosuvastatin, but not by atorvastatin


      • Platelet tissue factor (TF) initiates the clotting process.
      • TF activation is induced by VWF binding to GPIbα.
      • Platelet FXa generation in hypercholesterolemia (HC) is abnormally high.
      • Rosuvastatin decreases platelet cholesterol and TF-induced clotting in HC.
      • HDL and platelet TF activity correlate negatively in non-treated HC-patients.


      Background and aims

      High plasma LDL-cholesterol (LDL-C) and platelet responses have major pathogenic roles in atherothrombosis. Thus, statins and anti-platelet drugs constitute mainstays in cardiovascular prevention/treatment. However, the role of platelet tissue factor-dependent procoagulant activity (TF-PCA) has remained unexplored in hypercholesterolemia. We aimed to study platelet TF-PCA and its relationship with membrane cholesterol in vitro and in 45 hypercholesterolemic patients (HC-patients) (LDL-C >3.37 mmol/L, 130 mg/dL) and 37 control subjects (LDL-C <3.37 mmol/L). The effect of 1-month administration of 80 mg/day atorvastatin (n = 21) and 20 mg/day rosuvastatin (n = 24) was compared.


      Platelet TF-PCA was induced by GPIbα activation with VWF-ristocetin.


      Cholesterol-enriched platelets in vitro had augmented aggregation/secretion and platelet FXa generation (1.65-fold increase, p = 0.01). HC-patients had 1.5-, 2.3- and 2.5-fold increases in platelet cholesterol, TF protein and activity, respectively; their platelets had neither hyper-aggregation nor endogenous thrombin generation (ETP). Rosuvastatin, but not atorvastatin, normalized platelet cholesterol, TF protein and FXa generation. It also increased slightly the plasma HDL-C levels, which correlated negatively with TF-PCA.


      Platelets from HC-patients were not hyper-responsive to low concentrations of classical agonists and had normal PRP-ETP, before and after statin administration. However, washed platelets from HC-patients had increased membrane cholesterol, TF protein and TF-PCA. The platelet TF-dependent PCA was specifically expressed after VWF-induced GPIbα activation. Rosuvastatin, but not atorvastatin treatment, normalized the membrane cholesterol, TF protein and TF-PCA in HC-patients, possibly unveiling a new pleiotropic effect of rosuvastatin. Modulation of platelet TF-PCA may become a novel target to prevent/treat atherothrombosis without increasing bleeding risks.


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        • Taylor F.
        • Huffman M.D.
        • Macedo A.F.
        • et al.
        Statins for the primary prevention of cardiovascular disease.
        Cochrane database Syst. Rev. 2013; 1 (CD004816)
        • Gutierrez J.
        • Ramirez G.
        • Rundek T.
        • et al.
        Statin therapy in the prevention of recurrent cardiovascular events: a sex-based meta-analysis.
        Archives Intern. Med. 2012; 172: 909-919
        • Davi G.
        • Patrono C.
        Platelet activation and atherothrombosis.
        New England J. Med. 2007; 357: 2482-2494
        • Shattil S.J.
        • Anaya-Galindo R.
        • Bennett J.
        • et al.
        Platelet hypersensitivity induced by cholesterol incorporation.
        J. Clin. investigation. 1975; 55: 636-643
        • Shattil S.J.
        • Bennett J.S.
        • Colman R.W.
        • et al.
        Abnormalities of cholesterol-phospholipid composition in platelets and low-density lipoproteins of human hyperbetalipoproteinemia.
        J. laboratory Clin. Med. 1977; 89: 341-353
        • Lacoste L.
        • Lam J.Y.
        • Hung J.
        • et al.
        Hyperlipidemia and coronary disease. Correction of the increased thrombogenic potential with cholesterol reduction.
        Circulation. 1995; 92: 3172-3177
        • Owens 3rd, A.P.
        • Passam F.H.
        • Antoniak S.
        • et al.
        Monocyte tissue factor-dependent activation of coagulation in hypercholesterolemic mice and monkeys is inhibited by simvastatin.
        J. Clin. investigation. 2012; 122: 558-568
        • Narverud I.
        • Halvorsen B.
        • Nenseter M.S.
        • et al.
        Oxidized LDL level is related to gene expression of tumour necrosis factor super family members in children and young adults with familial hypercholesterolaemia.
        J. Intern. Med. 2013; 273: 69-78
        • Puccetti L.
        • Sawamura T.
        • Pasqui A.L.
        • et al.
        Atorvastatin reduces platelet-oxidized-LDL receptor expression in hypercholesterolaemic patients.
        Eur. J. Clin. investigation. 2005; 35: 47-51
        • Wraith K.S.
        • Magwenzi S.
        • Aburima A.
        • et al.
        Oxidized low-density lipoproteins induce rapid platelet activation and shape change through tyrosine kinase and Rho kinase-signaling pathways.
        Blood. 2013; 122: 580-589
        • Lafeber M.
        • Spiering W.
        • van der Graaf Y.
        • et al.
        The combined use of aspirin, a statin, and blood pressure-lowering agents (polypill components) and the risk of vascular morbidity and mortality in patients with coronary artery disease.
        Am. heart J. 2013; 166 (e281): 282-289
        • Hippisley-Cox J.
        • Coupland C.
        Effect of combinations of drugs on all cause mortality in patients with ischaemic heart disease: nested case-control analysis.
        BMJ Clin. Res. ed.). 2005; 330: 1059-1063
        • Luzak B.
        • Rywaniak J.
        • Stanczyk L.
        • et al.
        Pravastatin and simvastatin improves acetylsalicylic acid-mediated in vitro blood platelet inhibition.
        Eur. J. Clin. investigation. 2012; 42: 864-872
        • Malmstrom R.E.
        • Ostergren J.
        • Jorgensen L.
        • et al.
        Influence of statin treatment on platelet inhibition by clopidogrel - a randomized comparison of rosuvastatin, atorvastatin and simvastatin co-treatment.
        J. Intern. Med. 2009; 266: 457-466
        • Zhou Q.
        • Liao J.K.
        Statins and cardiovascular diseases: from cholesterol lowering to pleiotropy.
        Curr. Pharm. Des. 2009; 15: 467-478
        • Undas A.
        • Brummel-Ziedins K.E.
        • Mann K.G.
        Anticoagulant effects of statins and their clinical implications.
        Thrombosis haemostasis. 2014; 111: 392-400
        • Matarazzo S.
        • Quitadamo M.C.
        • Mango R.
        • et al.
        Cholesterol-lowering drugs inhibit lectin-like oxidized low-density lipoprotein-1 receptor function by membrane raft disruption.
        Mol. Pharmacol. 2012; 82: 246-254
        • Dietzen D.J.
        • Page K.L.
        • Tetzloff T.A.
        • et al.
        Inhibition of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase blunts factor VIIa/tissue factor and prothrombinase activities via effects on membrane phosphatidylserine, Arteriosclerosis.
        thrombosis, Vasc. Biol. 2007; 27: 690-696
        • Camera M.
        • Frigerio M.
        • Toschi V.
        • et al.
        Platelet activation induces cell-surface immunoreactive tissue factor expression, which is modulated differently by antiplatelet drugs, Arteriosclerosis.
        thrombosis, Vasc. Biol. 2003; 23: 1690-1696
        • Schwertz H.
        • Tolley N.D.
        • Foulks J.M.
        • et al.
        Signal-dependent splicing of tissue factor pre-mRNA modulates the thrombogenicity of human platelets.
        J. Exp. Med. 2006; 203: 2433-2440
        • Panes O.
        • Matus V.
        • Saez C.G.
        • et al.
        Human platelets synthesize and express functional tissue factor.
        Blood. 2007; 109: 5242-5250
        • Voight B.F.
        • Peloso G.M.
        • Orho-Melander M.
        • et al.
        Plasma HDL cholesterol and risk of myocardial infarction: a mendelian randomisation study.
        Lancet. 2012; 380: 572-580
        • Santos-Gallego C.G.
        • Badimon J.J.
        • Rosenson R.S.
        Beginning to understand high-density lipoproteins.
        Endocrinol. Metab. Clin. North Am. 2014; 43: 913-947
        • Briel M.
        • Ferreira-Gonzalez I.
        • You J.J.
        • et al.
        Association between change in high density lipoprotein cholesterol and cardiovascular disease morbidity and mortality: systematic review and meta-regression analysis.
        BMJ Clin. Res. ed.). 2009; 338 (b92)
        • Ridker P.M.
        • Genest J.
        • Boekholdt S.M.
        • et al.
        HDL cholesterol and residual risk of first cardiovascular events after treatment with potent statin therapy: an analysis from the JUPITER trial.
        Lancet. 2010; 376: 333-339
        • Quiroga T.
        • Goycoolea M.
        • Matus V.
        • et al.
        Diagnosis of mild platelet function disorders. Reliability and usefulness of light transmission platelet aggregation and serotonin secretion assays.
        Br. J. Haematol. 2009; 147: 729-736
        • Innis-Whitehouse W.
        • Li X.
        • Brown W.V.
        • et al.
        An efficient chromatographic system for lipoprotein fractionation using whole plasma.
        J. lipid Res. 1998; 39: 679-690
        • Rudel L.L.
        • Morris M.D.
        Determination of cholesterol using o-phthalaldehyde.
        J. lipid Res. 1973; 14: 364-366
        • Shankavaram U.T.
        • Lai W.C.
        • Netzel-Arnett S.
        • et al.
        Monocyte membrane type 1-matrix metalloproteinase. Prostaglandin-dependent regulation and role in metalloproteinase-2 activation.
        J. Biol. Chem. 2001; 276: 19027-19032
        • Hemker H.C.
        • Giesen P.L.
        • Ramjee M.
        • et al.
        The thrombogram: monitoring thrombin generation in platelet-rich plasma.
        Thrombosis haemostasis. 2000; 83: 589-591
        • Maurer E.
        • Tang C.
        • Schaff M.
        • et al.
        Targeting platelet GPIbbeta reduces platelet adhesion, GPIb signaling and thrombin generation and prevents arterial thrombosis, Arteriosclerosis.
        thrombosis, Vasc. Biol. 2013; 33: 1221-1229
        • Ravanat C.
        • Strassel C.
        • Hechler B.
        • et al.
        A central role of GPIb-IX in the procoagulant function of platelets that is independent of the 45-kDa GPIbalpha N-terminal extracellular domain.
        Blood. 2010; 116: 1157-1164
        • Cosemans J.M.
        • Schols S.E.
        • Stefanini L.
        • et al.
        Key role of glycoprotein Ib/V/IX and von Willebrand factor in platelet activation-dependent fibrin formation at low shear flow.
        Blood. 2011; 117: 651-660
        • van Lier M.
        • Verhoef S.
        • Cauwenberghs S.
        • et al.
        Role of membrane cholesterol in platelet calcium signalling in response to VWF and collagen under stasis and flow.
        Thrombosis haemostasis. 2008; 99: 1068-1078
        • Aoki I.
        • Aoki N.
        • Kawano K.
        • et al.
        Platelet-dependent thrombin generation in patients with hyperlipidemia.
        J. Am. Coll. Cardiol. 1997; 30: 91-96
        • Banfi C.
        • Brioschi M.
        • Lento S.
        • et al.
        Statins prevent tissue factor induction by protease-activated receptors 1 and 2 in human umbilical vein endothelial cells in vitro.
        J. thrombosis haemostasis JTH. 2011; 9: 1608-1619
        • Lindhout T.
        • Wielders S.
        • Hamulyak K.
        • et al.
        Inhibition of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase does not inhibit the platelet procoagulant response.
        J. thrombosis haemostasis JTH. 2008; 6: 1424-1426
        • Murrow J.R.
        • Sher S.
        • Ali S.
        • et al.
        The differential effect of statins on oxidative stress and endothelial function: atorvastatin versus pravastatin.
        J. Clin. Lipidol. 2012; 6: 42-49
        • Sironi L.
        • Gianazza E.
        • Gelosa P.
        • et al.
        Rosuvastatin, but not simvastatin, provides end-organ protection in stroke-prone rats by antiinflammatory effects, Arteriosclerosis.
        thrombosis, Vasc. Biol. 2005; 25: 598-603
        • Awasthi V.
        • Mandal S.K.
        • Papanna V.
        • et al.
        Modulation of tissue factor-factor VIIa signaling by lipid rafts and caveolae, Arteriosclerosis.
        thrombosis, Vasc. Biol. 2007; 27: 1447-1455
        • Munday A.D.
        • Gaus K.
        • Lopez J.A.
        The platelet glycoprotein Ib-IX-V complex anchors lipid rafts to the membrane skeleton: implications for activation-dependent cytoskeletal translocation of signaling molecules.
        J. thrombosis haemostasis JTH. 2010; 8: 163-172
        • Gousset K.
        • Wolkers W.F.
        • Tsvetkova N.M.
        • et al.
        Evidence for a physiological role for membrane rafts in human platelets.
        J. Cell. physiology. 2002; 190: 117-128
        • Pitt B.
        • Loscalzo J.
        • Monyak J.
        • et al.
        Comparison of lipid-modifying efficacy of rosuvastatin versus atorvastatin in patients with acute coronary syndrome (from the LUNAR study).
        Am. J. Cardiol. 2012; 109: 1239-1246
        • Liu B.
        • Cao H.M.
        • Li G.Y.
        • et al.
        Effects of rosuvastatin versus atorvastatin on rho-associated coiled-coil containing protein kinase activity and endothelial function in patients with atherosclerosis.
        J. Int. Med. Res. 2011; 39: 2314-2322
        • Tanaka S.
        • Fukumoto Y.
        • Nochioka K.
        • et al.
        Statins exert the pleiotropic effects through small GTP-binding protein dissociation stimulator upregulation with a resultant Rac1 degradation.
        Arteriosclerosis, thrombosis, Vasc. Biol. 2013; 33: 1591-1600
        • Badimon J.J.
        • Santos-Gallego C.G.
        • Badimon L.
        [Importance of HDL cholesterol in atherothrombosis: how did we get here? Where are we going?].
        Rev. Esp. Cardiol. 2010; 63: 20-35
        • Calkin A.C.
        • Drew B.G.
        • Ono A.
        • et al.
        Reconstituted high-density lipoprotein attenuates platelet function in individuals with type 2 diabetes mellitus by promoting cholesterol efflux.
        Circulation. 2009; 120: 2095-2104
        • Novotny W.F.
        • Girard T.J.
        • Miletich J.P.
        • et al.
        Purification and characterization of the lipoprotein-associated coagulation inhibitor from human plasma.
        J. Biol. Chem. 1989; 264: 18832-18837
        • Lesnik P.
        • Vonica A.
        • Guerin M.
        • et al.
        Anticoagulant activity of tissue factor pathway inhibitor in human plasma is preferentially associated with dense subspecies of LDL and HDL and with Lp(a).
        Arteriosclerosis thrombosis a J. Vasc. Biol./Am. Heart Assoc. 1993; 13: 1066-1075