Increased levels of platelet activation markers are positively associated with carotid wall thickness and other atherosclerotic risk factors in obese patients

Summary

The role of platelets in the development of atherosclerosis and obesity-related prothrombotic state is still under investigation. In this cross-sectional cohort study, we measured the levels of different platelet activation markers and evaluated their relationship with carotid intima-media thickness (IMT) along with other atherosclerotic risk factors in obese patients with or without atherosclerotic co-morbidities. We enrolled 154 obese patients, including 98 with either hypertension, type 2 diabetes mellitus or dyslipidaemia, 56 without these co-morbidities and 62 age- and sex-matched healthy controls. Platelet P-selectin expression and the number of platelet-derived microparticles (PMPs) were measured by flow cytometry; soluble P-selectin levels were analysed by ELISA and Thr715Pro P-selectin polymorphism was determined by PCR-RFLP. Carotid IMT was examined by ultrasonography. The levels of platelet activation parameters were significantly elevated in all obese subjects with increased carotid IMT compared to healthy controls. There was no effect of Thr715Pro genotype on soluble P-selectin levels in obese individuals contrary to normal subjects. Significant and positive association was revealed between carotid IMT and platelet P-selectin (p<0.0001), soluble P-selectin (p=0.039) and PMP (p=0.0001) levels. After adjusting for multiple variables, independent association was found between soluble P-selectin and fibrinogen (p=0.007), PMP levels and body mass index (p<0.0001) as well as platelet P-selectin and carotid IMT (p=0.012) plus plasminogen activator inhibitor-1 (p=0.009). In conclusion, P-selectin and PMP levels showed positive associations with abnormal carotid IMT and other risk factors in obesity suggesting a critical role of enhanced platelet reactivity in atherosclerotic wall alteration.

Keywords

Atherosclerosis - platelets - carotid intima-media thickness - P-selectin - microparticles - obesity - Thr715Pro polymorphism

• References

• 1 Blann AD, Nadar SK, Lip GY. The adhesion molecule P-selectin and cardiovascular disease. Eur Heart J 2003; 24: 2166-2179.
• 2 Vidal C, Spaulding C, Picard F. et al. Flow cytometry detection of platelet pro-coagulation activity and microparticles in patients with unstable angina treated by percutaneous coronary angioplasty and stent implantation. Thromb Haemost 2001; 86: 784-790.
• 3 Jennings LK. Mechanisms of platelet activation: need for new strategies to protect against platelet-mediated atherothrombosis. Thromb Haemost 2009; 102: 248-257.
• 4 Zeller JA, Tschoepe D, Kessler C. Circulating platelets show increased activation in patients with acute cerebral ischemia. Thromb Haemost 1999; 81: 373-377.
• 5 Shimizu M, Kohara S, Yamamoto M. et al. Significant relationship between platelet activation and intra-media thickness of the carotid artery in patients with ischemic cerebrovascular disease. Thromb Res 2006; 117: 647-652.
• 6 Ferroni P, Basili S, Falco A. et al. Platelet activation in type 2 diabetes mellitus. J Thromb Haemost 2004; 2: 1282-1291.
• 7 Nagy Jr B, Csongrádi E, Bhattoa HP. et al. Investigation of Thr715Pro P-selectin gene polymorphism and soluble P-selectin levels in type 2 diabetes mellitus. Thromb Haemost 2007; 98: 186-191.
• 8 Massberg S, Brand K, Grüner S. et al. A critical role of platelet adhesion in the initiation of atherosclerotic lesion formation. J Exp Med. 2002; 196: 887-896.
• 9 Burger PC, Wagner DD. Platelet P-selectin facilitates atherosclerotic lesion development. Blood 2003; 101: 2661-2666.
• 10 Preston RA, Coffey JO, Materson BJ. et al. Elevated platelet P-selectin expression and platelet activation in high risk patients with uncontrolled severe hypertension. Atherosclerosis 2007; 192: 148-154.
• 11 Undas A, Wiek I, Stêpien E. et al. Hyperglycemia is associated with enhanced thrombin formation, platelet activation, and fibrin clot resistance to lysis in patients with acute coronary syndrome. Diabetes Care 2008; 31: 1590-1595.
• 12 Ferroni P, Basili S, Davi G. Platelet activation, inflammatory mediators and hyper-cholesterolemia. Curr Vasc Pharmacol 2003; 1: 157-169.
• 13 Kappelmayer J, Nagy Jr B, Miszti-Blasius K. et al. The emerging value of P-selectin as a disease marker. Clin Chem Lab Med 2004; 42: 475-486.
• 14 Polgar J, Matuskova J, Wagner DD. The P-selectin, tissue factor, coagulation triad. J Thromb Haemost 2005; 3: 1590-1596.
• 15 George FD. Microparticles in vascular diseases. Thromb Res 2008; 122 (Suppl. 01) S55-59.
• 16 Davi G, Ferroni P. Microparticles in type 2 diabetes mellitus. J Thromb Haemost 2005; 3: 1166-1167.
• 17 De Pergola G, Pannacciulli N. Coagulation and fibrinolysis abnormalities in obesity. J Endocrinol Invest 2002; 25: 899-904.
• 18 Gallistl S, Sudi KM, Borkenstein M. et al. Correlation between cholesterol, soluble P-selectin, and D-dimer in obese children and adolescents. Blood Coagul Fibrinolysis 2000; 11: 755-760.
• 19 Basili S, Pacini G, Guagnano MT. et al. Insulin resistance as a determinant of platelet activation in obese women. J Am Coll Cardiol 2006; 48: 2531-2538.
• 20 De Pergola G, Pannacciulli N, Coviello M. et al. sP-selectin plasma levels in obesity: association with insulin resistance and related metabolic and prothrombotic factors. Nutr Metab Cardiovasc Dis 2008; 18: 227-232.
• 21 Goichot B, Grunebaum L, Desprez D. et al. Circulating procoagulant microparticles in obesity. Diabetes Metab 2006; 32: 82-85.
• 22 Murakami T, Horigome H, Tanaka K. et al. Impact of weight reduction on production of platelet-derived microparticles and fibrinolytic parameters in obesity. Thromb Res 2007; 119: 45-53.
• 23 Ueba T, Haze T, Sugiyama M. et al. Level, distribution and correlates of platelet-derived microparticles in healthy individuals with special reference to the metabolic syndrome. Thromb Haemost 2008; 100: 280-285.
• 24 Samocha-Bonet D, Justo D, Rogowski O. et al. Platelet counts and platelet activation markers in obese subjects. Mediators Inflamm 2008; 83: 4153-4158.
• 25 Barbaux SC, Blankenberg S, Rupprecht HJ. et al. Associations between P-selectin gene polymorphisms and soluble P-selectin levels and their relation to coronary artery disease. Atheroscler Thromb Vasc Biol 2001; 21: 1668-1673.
• 26 Miller MA, Kerry SM, Dong Y. et al. Associations between the Thr715Pro P-selectin gene polymorphism and soluble P-selectin in a multiethnic population in South London. Thromb Haemost 2004; 92: 1060-1065.
• 27 Reiner AP, Carlson CS, Thyagarajan B. et al. Soluble P-selectin, SELP polymorphisms, and atherosclerotic risk in European-American and African-African young adults: the Coronary Artery Risk Development in Young Adults (CARDIA) Study. Arterioscler Thromb Vasc Biol 2008; 28: 1549-1555.
• 28 Lorenz MW, Markus HS, Bots ML. et al. Prediction of clinical cardiovascular events with carotid intima-media thickness: a systematic review and meta-analysis. Circulation 2007; 115: 459-467.
• 29 Matthews DR, Hosker JP, Rudenski AS. et al. Homeostasis model assessment: insulin resistance and beta-cell function from fasting plasma glucose and insulin concentrations in man. Diabetologia 1985; 28: 412-419.
• 30 Dobiásová M.. Atherogenic index of plasma [log(triglycerides/HDL-cholesterol)]: theoretical and practical implications. Clin Chem 2004; 50: 1113-1115.
• 31 Nagy Jr B, Szuk T, Debreceni IB. et al. Platelet-derived microparticle levels are significantly elevated in patients treated by elective stenting compared to subjects with diagnostic catheterization alone. Platelets 2010; 21: 147-151.
• 32 MRC European Carotid Surgery Trial: interim results for symptomatic patients with severe (70–99%) or with mild (0–29%) carotid stenosis. European Carotid Surgery Trialists’ Collaborative Group. Lancet 1991; 337: 1235-1243.
• 33 Widder B, von Reutern GM, Neuerburg-Heusler D. Morphologic and Doppler sonographic criteria for determining the degree of stenosis of the internal carotid artery. Ultraschall Med 1986; 7: 70-75.
• 34 Howard G, Sharrett AR, Heiss G. et al. Carotid artery intimal-medial thickness distribution in general populations as evaluated by B-mode ultrasound. ARIC Investigators. Stroke 1993; 24: 1297-1304.
• 35 Lacoste L, Lam JY, Hung J. et al. Hyperlipidemia and coronary disease. Correction of the increased thrombogenic potential with cholesterol reduction. Circulation 1995; 92: 3172-3177.
• 36 Libby P. Inflammation in atherosclerosis. Nature 2002; 420: 868-874.
• 37 Ross R. Atherosclerosis – an inflammatory disease. N Engl J Med 1999; 340: 115-126.
• 38 Nagy Jr B, Jin J, Ashby B. et al. Contribution of the P2Y12 receptor-mediated pathway to platelet hyperreactivity in hypercholesterolemia. J Thromb Haemost 2011; 9: 810-819.
• 39 Koyama H, Maeno T, Fukumoto S. et al. Platelet P-selectin expression is associated with atherosclerotic wall thickness in carotid artery in humans. Circulation 2003; 108: 524-529.
• 40 Fusegawa Y, Hashizume H, Okumura T. et al. Hypertensive patients with carotid artery plaque exhibit increased platelet aggregability. Thromb Res 2006; 117: 615-622.
• 41 Lukasik M, Rozalski M, Luzak B. et al. Platelet activation and reactivity in the convalescent phase of ischaemic stroke. Thromb Haemost 2010; 103: 644-650.
• 42 Kappelmayer J, Berecki D, Misz M. et al. Monocytes express tissue factor in young patients with cerebral ischemia. Cerebrovasc Dis 1998; 8: 235-239.
• 43 Schneider DJ, Hardison RM, Lopes N. et al. Pro-Thrombosis Ancillary Study Group. Association between increased platelet P-selectin expression and obesity in patients with type 2 diabetes: a BARI 2D (Bypass Angioplasty Revascularization Investigation 2 Diabetes) substudy. Diabetes Care 2009; 32: 944-949.
• 44 Pannacciulli N, Cantatore FP, Minenna A. et al. C-reactive protein is independently associated with total body fat, central fat, and insulin resistance in adult women. Int J Obes Relat Metab Disord 2001; 25: 1416-1420.
• 45 Rosito GA, D'Agostino RB, Massaro J. et al. Association between obesity and a prothrombotic state: the Framingham Offspring Study. Thromb Haemost 2004; 91: 683-689.
• 46 Pannacciulli N, De Mitrio V, Marino R. et al. Effect of glucose tolerance status on PAI-1 plasma levels in overweight and obese subjects. Obes Res 2002; 10: 717-725.
• 47 Juhan-Vague I, Alessi MC, Mavri A. et al. Plasminogen activator inhibitor-1, inflammation, obesity, insulin resistance and vascular risk. J Thromb Haemost 2003; 1: 1575-1579.
• 48 Bucciarelli P, Sramek A, Reiber JH, Rosendaal FR. Arterial intima-media thickness and its relationship with cardiovascular disease and atherosclerosis: a possible contribution of medium-sized arteries. Thromb Haemost 2002; 88: 961-966.
• 49 Du XJ, Shan L, Gao XM, Kiriazis H, Liu Y, Lobo A, Head GA, Dart AM. Role of intramural platelet thrombus in the pathogenesis of wall rupture and intraventricular thrombosis following acute myocardial infarction. Thromb Haemost 2011; 105: 356-364.
• 50 Fateh-Moghadam S, Li Z, Ersel S. et al. Platelet degranulation is associated with progression of intima-media thickness of the common carotid artery in patients with diabetes mellitus type 2. Arterioscler Thromb Vasc Biol 2005; 25: 1299-1303.
• 51 Balla J, Magyar MT, Bereczki D. et al. Serum levels of platelet released CD40 ligand are increased in early onset occlusive carotid artery disease. Dis Markers 2006; 22: 133-140.
• 52 Volcik KA, Ballantyne CM, Coresh J. et al. P-selectin Thr715Pro polymorphism predicts P-selectin levels but not risk of incident coronary heart disease or ischemic stroke in a cohort of 14595 participants: The Atherosclerosis Risk In Communities Study. Atherosclerosis 2006; 186: 74-79.