Atorvastatin reduces thrombin generation and expression of tissue factor, P-selectin and GPIIIa on platelet-derived microparticles in patients with peripheral arterial occlusive disease

Fariborz Mobarrez; Shu He; Anders Bröijersen; Björn Wiklund; Aleksandra Antovic; Jovan Antovic; Nils Egberg; Gun Jörneskog; Håkan Wallén

doi:10.1160/TH10-12-0810

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2011; 106(02): 344-352
DOI: 10.1160/TH10-12-0810

Platelets and Blood Cells

Schattauer GmbH

Atorvastatin reduces thrombin generation and expression of tissue factor, P-selectin and GPIIIa on platelet-derived microparticles in patients with peripheral arterial occlusive disease

Fariborz Mobarrez

¹Karolinska Institutet Department of Clinical Sciences, Danderyd Hospital, Division of Cardiovascular Medicine, Stockholm, Sweden

,

Shu He

²Coagulation Research, Department of Molecular Medicine & Surgery, Karolinska Institutet & Clinical Chemistry, Karolinska University Hospital, Solna, Sweden

,

Anders Bröijersen

³Department of Medicine, Karolinska University Hospital, Huddinge, Sweden

,

Björn Wiklund

⁴Department of Vascular Surgery, Karolinska University Hospital, Huddinge, Sweden

,

Aleksandra Antovic

¹Karolinska Institutet Department of Clinical Sciences, Danderyd Hospital, Division of Cardiovascular Medicine, Stockholm, Sweden

,

Jovan Antovic

²Coagulation Research, Department of Molecular Medicine & Surgery, Karolinska Institutet & Clinical Chemistry, Karolinska University Hospital, Solna, Sweden

,

Nils Egberg

²Coagulation Research, Department of Molecular Medicine & Surgery, Karolinska Institutet & Clinical Chemistry, Karolinska University Hospital, Solna, Sweden

,

Gun Jörneskog

⁵KarolinskaInstitutet, Department of Clinical Sciences, Danderyd Hospital, Division of Internal Medicine, Stockholm, Sweden

,

Håkan Wallén

¹Karolinska Institutet Department of Clinical Sciences, Danderyd Hospital, Division of Cardiovascular Medicine, Stockholm, Sweden

› Author Affiliations

Abstract

Summary

We investigated the effects of statin treatment on platelet-derived microparticles (PMPs) and thrombin generation in atherothrombotic disease. Nineteen patients with peripheral arterial occlusive disease were randomised to eight weeks of treatment with atorvastatin or placebo in a cross-over fashion. Expression of GPIIIa (CD61), P-selectin (CD62P), tissue factor (TF, CD142) and phosphatidylserine (PS; annexin-V or lactadherin binding) was assessed on PMPs. Thrombin generation in vivo was assessed by measurement of prothrombin fragment 1+2 in plasma (F1+2) and ex vivo by using the calibrated automated thrombogram (CAT). During atorvastatin treatment, expression of TF, P-selectin and GPIIIa was significantly reduced vs. placebo (p<0.001 for all). No effect on annexin-V or lactadherin binding was seen. Thrombin generation was significantly reduced during atorvastatin as assessed by both the CAT assay (p<0.001) and by measurements of F1+2 (p<0.01). Subsequent in vitro experiments showed that when TF on microparticles (MPs) was blocked by antibodies, the initiation of thrombin generation was slightly but significantly delayed. Blocking PS on MPs using annexin-V or lactadherin resulted in almost complete inhibition of thrombin generation. In conclusion, atorvastatin reduces thrombin generation and expression of TF, GPIIIa and P-selectin on PMPs in patients with peripheral vascular disease. Microparticle-bound TF slightly enhances initiation of thrombin generation whereas negatively charged surfaces provided by MPs or lipoproteins could reinforce thrombin generation. Statins may inhibit initiation of thrombin generation partly through a microparticle dependent mechanism but the main effect is probably through reduction of lipoprotein levels.

Keywords

Atorvastatin - peripheral arterial occlusive disease - platelet-derived microparticles - thrombin generation - tissue factor

Full Text

References

References
1 Hooi JD, Kester AD, Stoffers HE. et al. Asymptomatic peripheral arterial occlusive disease predicted cardiovascular morbidity and mortality in a 7-year follow-up study. J Clin Epidemiol 2004; 57: 294-300.
2 Hackam DG, Eikelboom JW. Antithrombotic treatment for peripheral arterial disease. Heart 2007; 93: 303-308.
3 Ludman A, Venugopal V, Yellon DM. et al. Statins and cardioprotection–more than just lipid lowering?. Pharmacol Ther 2009; 122: 30-43.
4 Momsen AH, Jensen MB, Norager CB. et al. Drug therapy for improving walking distance in intermittent claudication: a systematic review and meta-analysis of robust randomised controlled studies. Eur J Vasc Endovasc Surg 2009; 38: 463-474.
5 Ray KK, Cannon CP. The potential relevance of the multiple lipid-independent (pleiotropic) effects of statins in the management of acute coronary syndromes. J Am Coll Cardiol 2005; 46: 1425-1433.
6 George FD. Microparticles in vascular diseases. Thromb Res. 2008; 122 (Suppl. 01) S55-59
7 Boulanger CM. Microparticles, vascular function and hypertension. Curr Opin Nephrol Hypertens. 2010; 19: 177-180.
8 Morel O, Toti F, Hugel B. et al. Procoagulant microparticles: disrupting the vascular homeostasis equation?. Arterioscler Thromb Vasc Biol 2006; 26: 2594-2604.
9 Morel O, Toti F, Hugel B. et al. Cellular microparticles: a disseminated storage pool of bioactive vascular effectors. Curr Opin Hematol 2004; 11: 156-164.
10 Burnier L, Fontana P, Kwak BR. et al. Cell-derived microparticles in haemostasis and vascular medicine. Thromb Haemost 2009; 101: 439-451.
11 Yuana Y, Bertina RM, Osanto S. Pre-analytical and analytical issues in the analysis of blood microparticles. Thromb Haemost 2010; 105: 396-408.
12 Hoffman M, Monroe DM. 3rd. A cell-based model of hemostasis. Thromb Haemost 2001; 85: 958-965.
13 Perneby C, Granström E, Beck O. et al. Optimization of an enzyme immunoassay for 11-dehydro-thromboxane B(2) in urine: comparison with GC-MS. Thromb Res 1999; 96: 427-436.
14 Hemker HC, Al Dieri R, De Smedt E. et al. Thrombin generation, a function test of the haemostatic-thrombotic system. Thromb Haemost 2006; 96: 553-561.
15 Mobarrez F, Antovic J, Egberg N. et al. A multicolor flow cytometric assay for measurement of platelet-derived microparticles. Thromb Res 2010; 125: e110-116.
16 Dasgupta SK, Abdel-Monem H, Niravath P. et al. Lactadherin and clearance of platelet-derived microvesicles. Blood 2009; 113: 1332-1339.
17 Takemoto M, Liao JK. Pleiotropic effects of 3-hydroxy-3-methylglutaryl coenzyme a reductase inhibitors. Arterioscler Thromb Vasc Biol 2001; 21: 1712-1719.
18 Labiós M, Martínez M, Gabriel F. et al. Effect of atorvastatin upon platelet activation in hypercholesterolemia, evaluated by flow cytometry. Thromb Res 2005; 115: 263-270.
19 Malmström RE, Settergren M, Böhm F. et al. No effect of lipid lowering on platelet activity in patients with coronary artery disease and type 2 diabetes or impaired glucose tolerance. Thromb Haemost 2009; 101: 157-164.
20 Lijnen P, Echevaría-Vázquez D, Petrov V. Influence of cholesterol-lowering on plasma membrane lipids and function. Methods Find Exp Clin Pharmacol 1996; 18: 123-136.
21 Vaughan CJ, Gotto AM, Jr Basson CT. The evolving role of statins in the management of atherosclerosis. J Am Coll Cardiol 2000; 35: 1-10.
22 Bach RR. Tissue factor encryption. Arterioscler Thromb Vasc Biol 2006; 26: 456-461.
23 Tramontano AF, O’Leary J, Black AD. et al. Statin decreases endothelial microparticle release from human coronary artery endothelial cells: implication for the Rho-kinase pathway. Biochem Biophys Res Commun 2004; 320: 34-38.
24 Sommeijer DW, Joop K, Leyte A. et al. Pravastatin reduces fibrinogen receptor gpIIIa on platelet-derived microparticles in patients with type 2 diabetes. J Thromb Haemost 2005; 3: 1168-1171.
25 Nomura S, Shouzu A, Omoto S. et al. Effects of losartan and simvastatin on monocyte-derived microparticles in hypertensive patients with and without type 2 diabetes mellitus. Clin Appl Thromb Hemost 2004; 10: 133-141.
26 Tehrani S, Mobarrez F, Antovic A. et al. Atorvastatin has antithrombotic effects in patients with type 1 diabetes and dyslipidemia. Thromb Res 2010; 126: e225-31.
27 Lechner D, Weltermann A. Circulating tissue factor-exposing microparticles. Thromb Res 2008; 122: 47-54.
28 Diamant M, Nieuwland R, Pablo RF. et al. Elevated numbers of tissue-factor exposing microparticles correlate with components of the metabolic syndrome in uncomplicated type 2 diabetes mellitus. Circulation 2002; 106: 2442-2447.
29 Rota S, William NA, Baglin TP. et al. Atherogenic lipoproteins support assembly of the prothrombinase complex and thrombin generation: modulation by oxidation and vitamin E. Blood 1999; 91: 508-515.
30 Bröijersén A, Hamsten A, Silveira A. et al. Gemfibrozil reduces thrombin generation in patients with combined hyperlipidaemia, without influencing plasma fibrinogen, fibrin gel structure or coagulation factor VII. Thromb Haemost 1996; 76: 171-176.
31 Notarbartolo A, Davì G, Averna M. et al. Inhibition of thromboxane biosynthesis and platelet function by simvastatin in type IIa hypercholesterolemia. Arterioscler Thromb Vasc Biol 1995; 15: 247-251.