Oxidative stress and platelet activation in subjects with moderate hyperhomocysteinaemia due to MTHFR 677 C→T polymorphism

Alfredo Dragani; Angela Falco; Francesca Santilli; Stefania Basili; Giancarlo Rolandi; Loredana Cerasa; Stefano Lattanzio; Giovanni Ciabattoni; Carlo Patrono; Giovanni Davì

doi:10.1160/TH11-12-0899

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2012; 108(03): 533-542
DOI: 10.1160/TH11-12-0899

Platelets and Blood Cells

Schattauer GmbH

Oxidative stress and platelet activation in subjects with moderate hyperhomocysteinaemia due to MTHFR 677 C→T polymorphism

Autoren

Alfredo Dragani

¹Departments of Haematology and Laboratory Medicine, Civil Hospital Pescara, Italy
Angela Falco

²“G. d’Annunzio” University Foundation and Department of Drug Sciences, University of Chieti “G. d’Annunzio” Schools of Medicine and Pharmacy, Chieti, Italy
Francesca Santilli

²“G. d’Annunzio” University Foundation and Department of Drug Sciences, University of Chieti “G. d’Annunzio” Schools of Medicine and Pharmacy, Chieti, Italy
Stefania Basili

³Department of Medicine, “La Sapienza”, University of Rome, Rome, Italy
Giancarlo Rolandi

¹Departments of Haematology and Laboratory Medicine, Civil Hospital Pescara, Italy
Loredana Cerasa

²“G. d’Annunzio” University Foundation and Department of Drug Sciences, University of Chieti “G. d’Annunzio” Schools of Medicine and Pharmacy, Chieti, Italy
Stefano Lattanzio

²“G. d’Annunzio” University Foundation and Department of Drug Sciences, University of Chieti “G. d’Annunzio” Schools of Medicine and Pharmacy, Chieti, Italy
Giovanni Ciabattoni

²“G. d’Annunzio” University Foundation and Department of Drug Sciences, University of Chieti “G. d’Annunzio” Schools of Medicine and Pharmacy, Chieti, Italy
Carlo Patrono

⁴Department of Pharmacology, Catholic University School of Medicine, Rome, Italy
Giovanni Davì

²“G. d’Annunzio” University Foundation and Department of Drug Sciences, University of Chieti “G. d’Annunzio” Schools of Medicine and Pharmacy, Chieti, Italy

Abstract

Summary

The methylenetetrahydrofolate reductase (MTHFR) 677 C→T polymorphism may be associated with elevated total homocysteine (tHcy) levels, an independent risk factor for cardiovascular disease. It was the study objective to evaluate in vivo lipid peroxidation and platelet activation in carriers of the MTHFR 677 C→T polymorphism and in non-carriers, in relation to tHcy and folate levels. A cross-sectional comparison of urinary 8-iso-prostaglandin (PG)F_2α and 11-dehydro-thromboxane (TX)B₂ (markers of in vivo lipid peroxidation and platelet activation, respectively) was performed in 100 carriers and 100 non-carriers of the polymorphism. A methionine-loading test and folic acid supplementation were performed to investigate the causal relationship of the observed associations. Urinary 8-iso-PGF_2α and 11-dehydro-TXB₂ were higher in carriers with hyperhomocysteinaemia than in those without hyperhomocysteinaemia (p<0.0001). Hyperhomocysteinaemic carriers had lower folate levels (p=0.0006), higher urinary 8-iso-PGF_2α (p<0.0001) and 11-dehydro-TXB₂ (p<0.0001) than hyperhomocysteinaemic non-carriers. On multiple regression analysis, high tHcy (p<0.0001), low folate (p<0.04) and MTHFR 677 C→T polymorphism (p<0.001) independently predicted high rates of 8-iso-PGF_2α excretion. Methionine loading increased plasma tHcy (p=0.002), and both urinary prostanoid metabolites (p=0.002). Folic acid supplementation was associated with decreased urinary 8-iso-PGF_2α and 11-dehydro-TXB₂ excretion (p<0.0003) in the hyperhomocysteinaemic group, but not in the control group, with substantial inter-individual variability related to baseline tHcy level and the extent of its reduction. In conclusion, hyperhomocysteinaemia due to the MTHFR 677 C→T polymorphism is associated with enhanced in vivo lipid peroxidation and platelet activation that are reversible, at least in part, following folic acid supplementation. An integrated biomarker approach may help identifying appropriate candidates for effective folate supplementation.

Keywords

Hyperhomocysteinaemia - MTHFR 677 C→T polymorphism - oxidative stress - platelet activation - folate supplementation

Volltext

Referenzen

References
1 Welch GN, Loscalzo J. Homocysteine and atherothrombosis. N Engl J Med 1998; 338: 1042-1050.
2 Durand P. et al. Impaired homocysteine metabolism and atherothrombotic disease. Lab Invest 2001; 81: 645-672.
3 Wald DS. et al. Homocysteine and cardiovascular disease: evidence on causality from a meta-analysis. Br Med J 2002; 325: 1202-1208.
4 The Homocysteine Studies Collaboration. Homocysteine and risk of ischemic heart disease and stroke: a meta-analysis. J Am Med Assoc. 2002 288. 2015-2022.
5 Klerk M. et al. MTHFR Studies Collaboration Group. MTHFR 677C→T polymorphism and risk of coronary heart disease: a meta-analysis. J Am Med Assoc 2002; 288: 2023-2031.
6 Zee RY. et al. Homocysteine, 5,10-methylenetetrahydrofolate reductase 677C>T polymorphism, nutrient intake, and incident cardiovascular disease in 24,968 initially healthy women. Clin Chem 2007; 53: 845-851.
7 Lewis SJ. et al. Meta-analysis of MTHFR 677C->T polymorphism and coronary heart disease: does totality of evidence support causal role for homocysteine and preventive potential of folate?. Br Med J 2005; 331: 1053.
8 Malinow MR. et al. The effects of folic acid supplementation on plasma total homocysteine are modulated by multivitamin use and methylenetetrahydrofolate reductase genotypes. Arterioscler Thromb Vasc Biol 1997; 17: 1157-1162.
9 Holmes M V. et al. Effect modification by population dietary folate on the association between MTHFR genotype, homocysteine, and stroke risk: a meta-analysis of genetic studies and randomised trials. Lancet 2011; 378: 584-594.
10 Liem A. et al. Secondary prevention with folic acid: effects on clinical outcomes. J Am Coll Cardiol 2003; 41: 2105-2113.
11 Bonaa KH. et al. NORVIT Trial Investigators. Homocysteine lowering and cardiovascular events after acute myocardial infarction. N Engl J Med 2006; 354: 1578-1588.
12 Lonn E. et al. Heart Outcomes Prevention Evaluation (HOPE) 2 Investigators. Homocysteine lowering with folic acid and B vitamins in vascular disease. N Engl J Med 2006; 354: 1567-1577.
13 Albert CM. et al. Effect of Folic Acid and B Vitamins on Risk of Cardiovascular Events and Total Mortality Among Women at High Risk for Cardiovascular Disease: A Randomized Trial. J Am Med Assoc 2008; 299: 2027-2036.
14 SEARCH Collaborative Group. Effects of homocysteine-lowering with folic acid plus vitamin B12 vs placebo on mortality and major morbidity in myocardial infarction survivors: a randomized trial. J Am Med Assoc. 2010 303. 2486-2494.
15 Clarke R. et al. Effects of lowering homocysteine levels with B vitamins on cardiovascular disease, cancer, and cause-specific mortality: Meta-analysis of 8 randomized trials involving 37 485 individuals. Arch Intern Med 2010; 170: 1622-1631.
16 Clarke R. et al. Homocysteine and coronary heart disease: meta-analysis of MTHFR case-control studies, avoiding publication bias. PLoS Med 2012; 09: e1001177.
17 Loscalzo J. The oxidant stress of hyperhomocyst(e)inemia. J Clin Invest 2006; 98: 5-7.
18 Davì G. et al. Oxidative stress and platelet activation in homozygous homocystinuria. Circulation 2001; 104: 1124-1128.
19 Papatheodorou L, Weiss N. Vascular oxidant stress and inflammation in hyperhomocysteinemia. Antiox Redox Signal 2007; 09: 1941-1958.
20 Podrez EA. et al. Platelet CD36 links hyperlipidemia, oxidant stress and a prothrombotic phenotype. Nat Med 2007; 13: 1086-1095.
21 Patrono C, FitzGerald GA. Isoprostanes: potential markers of oxidant stress in atherothrombotic disease. Arterioscler Thromb Vasc Biol 1997; 17: 2309-2315.
22 Milne GL. et al. Human biochemistry of the isoprostane pathway. J Biol Chem 2008; 283: 15533-15537.
23 Braunwald E. Biomarkers in heart failure. N Engl J Med 2008; 358: 2148-2159.
24 Davì G, Patrono C. Platelet activation and atherothrombosis. N Engl J Med 2007; 357: 2482-2494.
25 Ciabattoni G. et al. Fractional conversion of thromboxane B₂ to urinary 11-dehydro-thromboxane B₂ in man. Biochim Biophys Acta 1989; 992: 66-70.
26 Frosst P. et al. A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet 1995; 10: 111-113.
27 Margaglione M. et al. The methylenetetrahydrofolate reductase TT677 genotype is associated with venous thrombosis independently of the coexistence of the FV Leiden and the prothrombin A20210 mutation. Thromb Haemost 1998; 79: 907-911.
28 Shipchandler MT, Moore EG. Rapid, fully automatic measurement of plasma homocyst(e)ine with the Abbott IMx analyzer. Clin Chem 1995; 41: 991-994.
29 Zighetti ML. et al. Determination of total homocysteine in plasma: comparison of the Abbott IMx immunoassay with high performance liquid chromatography. Haematologica 2002; 87: 89-94.
30 Wang Z. et al. Immunological characterization of urinary 8-epi-prostaglandin F_?excretion in man. J Pharmacol Exp Ther 1995; 275: 94-100.
31 Ciabattoni G. et al. Radioimmunoassay of 11-dehydro-TXB₂ in human plasma and urine. Biochim Biophys Acta 1987; 918: 293-297.
32 Martinelli I. et al. Hyperhomocysteinemia in cerebral vein thrombosis. Blood 2003; 102: 1363-1366.
33 Cattaneo M. Hyperhomocysteinemia, atherosclerosis and thrombosis. Thromb Haemost 1999; 81: 165-176.
34 Voutilainen S. et al. Enhanced in vivo lipid peroxidation at elevated plasma total homocysteine levels. Arterioscler Thromb Vasc Biol 1999; 19: 1263-1266.
35 Audoly LP. et al. Cardiovascular responses to the isoprostanes iPF(2alpha)-III and iPE(2)-III are mediated via the thromboxane A(2) receptor in vivo. Circulation 2000; 101: 2833-2840.
36 Minuz P. et al. The F2-isoprostane 8-epiprostaglandin F2alpha increases platelet adhesion and reduces the antiadhesive and antiaggregatory effects of NO. Arterioscler Thromb Vasc Biol 1998; 18: 1248-1256.
37 Pignatelli P. et al. Inherited human gp91phox deficiency is associated with impaired isoprostane formation and platelet dysfunction. Arterioscler Thromb Vasc Biol 2011; 31: 423-434.
38 Cronin S. et al. Dose-related association of MTHFR 677T allele with risk of ischemic stroke. Evidence from a cumulative meta-analysis. Stroke 2005; 36: 1581-1587.
39 Shirodaria C. et al. Global improvement of vascular function and redox state with low-dose folic acid: implications for folate therapy in patients with coronary artery disease. Circulation 2007; 115: 2262-2270.
40 Schnyder G. et al. Decreased rate of coronary restenosis after lowering of plasma homocysteine levels. N Engl J Med 2001; 345: 1593-1600.
41 Loscalzo J. Homocysteine trials – Clear outcomes for complex reasons. N Engl J Med 2006; 354: 1629-1632.
42 Wang X. et al. Efficacy of folic acid supplementation in stroke prevention: a meta-analysis. Lancet 2007; 369: 1876-1882.
43 Bazzano LA. et al. Effect of folic acid supplementation on risk of cardiovascular diseases: a meta-analysis of randomized controlled trials. J Am Med Assoc 2006; 296: 2720-2726.

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