RAS and MTHFR Gene Polymorphisms in a Healthy Exercise-trained Population: Association with the MTHFR (TT) Genotype and a Lower Hemoglobin Level

Abstract

The aim of this study was to determine the frequencies of ACE (I/D), AGT (M235T), AT1R (A1166C) and MTHFR (C677T) polymorphisms in a well-defined (in regards to health and nutritional status and lifestyle) population of young, healthy, exercise-trained subjects (no. 100) from the Campania region of Southern Italy. We also investigated whether there was any correlation between these polymorphisms and biochemical, hematological and hemostatic parameters in this “low-risk” population. Gene polymorphisms were analyzed with the polymerase chain reaction and restriction enzyme analysis. Allele frequencies of the genotypes examined were in Hardy-Weinberg equilibrium and agree with those reported in the Italian population. No associations were found between ACE, AGT, AT1R gene polymorphisms and anthropometric, clinical and laboratory parameters. However, the MTHFR (C677T) polymorphism was significantly associated with lower hemoglobin plasma levels in TT vs. CC + CT females (p < 0.016). This report is the first to describe the frequencies of RAS and MTHFR gene polymorphisms in young, exercise-trained volunteers from Campania and to identify an association between the MTHFR gene polymorphisms and lower hemoglobin plasma levels in young healthy females.

Key words

RAS and MTHFR gene polymorphisms - homocysteine level - hemoglobin level

References

• 1 Alvarez R, Terrados N, Ortolano R, Iglesias-Cubero G, Reguero J R, Batalla A, Cortina A, Fernandez-Garcia B, Rodriguez C, Braga S, Alvarez V, Coto E. Genetic variation in the renin-angiotensin system and athletic performance.  Eur J Appl Physiol. 2000;  82 117-120
• 2 Amant C, Hamon M, Bauters C, Richard F, Helbecque N, McFadden E P, Escudero X, Lablanche J M, Amouyel P, Bertrand M E. The angiotensin II type 1 receptor gene polymorphism is associated with coronary artery vasoconstriction.  J Am Coll Cardiol. 1997;  29 486-490
• 3 Arant C B, Wessel T R, Olson M B, Bairey Merz C N, Sopko G, Rogers W J, Sharaf B L, Reis S E, Smith K M, Johnson B D, Handberg E, Mankad S, Pepine C J. Hemoglobin level is an independent predictor for adverse cardiovascular outcomes in women undergoing evaluation for chest pain: results from the National Heart, Lung, and Blood Institute Women's Ischemia Syndrome Evaluation Study.  J Am Coll Cardiol. 2004;  43 2009-2014
• 4 Arca M, Pannitteri G, Campagna F, Candeloro A, Montali A, Cantini R, Seccareccia F, Campa P P, Marino B, Ricci G. Angiotensin-converting enzyme gene polymorphism is not assocciated with coronary atherosclerosis and myocardial infarction in a sample of Italian patients.  Eur J Clin Invest. 1998;  28 485-490
• 5 Botto L D, Yang Q. 5,10-Methylenetetrahydrofolate reductase gene variants and congenital anomalies: a HuGE review.  Am J Epidemiol. 2000;  151 862-877
• 6 Brattstrom L, Wilcken D E, Ohrvik J, Brudin L. Common methylenetetrahydrofolate reductase gene mutation leads to hyperhomocysteinemia but not to vascular disease: the result of a meta-analysis.  Circul. 1998;  98 2520-2526
• 7 Bray M S. Genomics, genes, and environmental interaction: the role of exercise.  J Appl Physiol. 2000;  88 788-792
• 8 Cambien F, Poirier O, Lecert L, Evans A, Cambou J P, Arveiler D, Luc G, Bard J M, Bara L, Ricard S, Tiret L, Amouyel P, Alhenc-Gelas F, Soubrier F. Deletion polymorphism in the gene for angiotensin-converting enzyme is a potent risk factor for myocardial infarction.  Nature. 1992;  359 641-644
• 9 Carluccio M, Soccio M, De Caterina R. Aspects of gene polymorphisms in cardiovascular disease: the renin-angiotensin system.  Eur J Clin Invest. 2001;  31 476-478
• 10 Celentano A, Mancini F P, Crivaro M, Palmieri V, De Stefano V, Ferrara L A, Di Minno G, de Simone G. Influence of cardiovascular risk factors on relation between angiotensin converting enzyme-gene polymorphism and blood pressure in arterial hypertension.  J Hypertens. 1998;  16 985-991
• 11 Chao-Hung Ho. The influence of age, sex, vitamin B12, folate levels and methylene-tetrahydrofolate reductase C677T genetic mutations on plasma homocysteine in the Chinese population.  Haematologica. 2000;  85 1051-1054
• 12 de Franchis R, Mancini F P, D'Angelo A, Sebastio G, Fermo I, de Stefano V, Margaglione M, Mazzola G, di Minno G, Andria G. Elevated total plasma homocysteine and 677C→T mutation of the 5, 10-methylenetetrahydrofolate reductase gene in thrombotic vascular disease.  Am J Hum Genet. 1996;  59 262-264
• 13 Diet F, Graf C, Mahnke N, Wassmer G, Predel H G, Palma-Hohmann I, Rost R, Bohm M. ACE and angiotensinogen gene genotypes and left ventricular mass in athletes.  Eur J Clin Invest. 2001;  31 836-842
• 14 Fatini C, Abbate R, Pepe G, Battaglini B, Gensini F, Ruggiano G, Gensini G F, Guazzelli R. Searching for a better assessment of the individual coronary risk profile. The role of angiotensin-converting enzyme, angiotensin II type 1 receptor and angiotensinogen gene polymorphisms.  Eur Heart J. 2000;  21 633-638
• 15 Fatini C, Guazzelli R, Manetti T, Battaglini B, Gensini F, Vono R, Toncelli L, Zilli P, Capalbo A, Abbate R, Gensini G F, Galanti G. RAS genes influence exercise-induced left ventricular hypertrophy: an elite-athletes study.  Med Sci Sports Exerc. 2000;  32 1868-1872
• 16 Fatini C, Gensini F, Sticchi E, Battaglini B, Prisco D, Fedi S, Brunelli T, Marcucci R, Conti A A, Gensini G F, Abbate R. ACE DD genotype: an independent predisposition factor to venous thromboembolism.  Eur J Clin Invest. 2003;  33 642-647
• 17 Friso S, Choi S W, Girelli D, Mason J B, Dolnikowski G G, Bagley P J, Olivieri O, Jacques P F, Rosenberg I H, Corrocher R, Selhub J. A common mutation in the 5,10-methylenetetrahydrofolate reductase gene affects genomic DNA methylation through an interaction with folate status.  Proc Natl Acad Sci USA. 2002;  99 5606-5611
• 18 Gayagay G, Yu B, Hambly B, Boston T, Hahn A, Celermajer D S, Trent R J. Elite endurance athletes and ACE I allele - the role of genes in athletic performance.  Hum Genet. 1998;  103 48-50
• 19 Garcia-Garcia I, Garcia-Fragoso L, Renta J, Arce S, Cadilla C L. Erythrocyte volume, folate levels, and the presence of methylenetetrahydrofolate reductase polymorphism.  P R Health Sci J. 2002;  21 17-19
• 20 Gardemann A, Nguyen Q D, Humme J, Stricker J, Katz N, Tillmanns H, Hehrlein F W, Rau M, Haberbosch W. Angiotensin II type 1 receptor A1166C gene polymorphism. Absence of an association with the risk of coronary artery disease and myocardial infarction and of a synergistic effect with angiotensin-converting enzyme gene polymorphism on the risk of these diseases.  Eur Heart J. 1998;  19 1657-1665
• 21 Geisel J, Hennen B, Hubner U, Knapp J P, Herrmann W. The impact of hyperhomocysteinemia as a cardiovascular risk factor in the prediction of coronary heart disease.  Clin Chem Lab Med. 2003;  41 1513-1517
• 22 Gonzalez Ordonez A J, Fernandez Carreira J M, Medina Rodriguez J M, Martin Sanchez L, Alvarez Diaz R, Alvarez Martinez M V, Coto Garcia E. Risk of venous thromboembolis associated with the insertion/deletion polymorphism in the angiotensin-converting enzyme gene.  Blood Coagul Fibrinolysis. 2001;  11 485-490
• 23 Guttormsen A B, Ueland P M, Nesthus I, Nygard O, Schneede J, Vollset S E, Refsum H. Determinants and vitamin responsiveness of intermediate hyperhomocysteinemia (> or = 40 micromol/liter). The Hordaland Homocysteine Study.  J Clin Invest. 1996;  98 2174-2183
• 24 Herrmann M, Wilkinson J, Schorr H, Obeid R, Georg T, Urhausen A, Scharhag J, Kindermann W, Herrmann W. Comparison of the influence of volume-oriented training and high-intensity interval training on serum homocysteine and its cofactors in young, healthy swimmers.  Clin Chem Lab Med. 2003;  41 1525-1531
• 25 Herrmann M, Schorr H, Obeid R, Scharhag J, Urhausen A, Kindermann W, Herrmann W. Homocysteine increases during endurance exercise.  Clin Chem Lab Med. 2003;  41 1518-1524
• 26 Hingorani A D, Brown M J. A simple molecular assay for the C1166 variant of the angiotensin II type 1 receptor gene.  Biochem Biophys Res Commun. 1995;  213 725-729
• 27 Hustad S, Ueland P M, Vollset S E, Zhang Y, Bjorke-Monsen A L, Schneede J. Riboflavin as a determinant of plasma total homocysteine: effect modification by the methylenetetrahydrofolate reductase C677T polymorphism.  Clin Chem. 2000;  46 1065-1071
• 28 Katsuya T, Koike G, Yee T W, Sharpe N, Jackson R, Norton R, Horiuchi M, Pratt R E, Dzau V J, MacMahon S. Association of angiotensinogen gene T235 variant with increased risk of coronary heart disease.  Lancet. 1995;  345 1660-1663
• 29 Keavney B, McKenzie C, Parish S, Palmer A, Clark S, Youngman L, Delepine M, Lathrop M, Peto R, Collins R. Large-scale test of hypothesised associations between the angiotensin-converting-enzyme insertion/deletion polymorphism and myocardial infarction in about 5000 cases and 6000 controls. International Studies of Infarct Survival (ISIS) Collaborators.  Lancet. 2000;  355 434-442
• 30 Kluijtmans L A, Young I S, Boreham C A, Murray L, McMaster D, McNulty H, Strain J J, McPartlin J, Scott J M, Whitehead A S. Genetic and nutritional factors contributing to hyperhomocysteinemia in young adults.  Blood. 2003;  101 2483-2488
• 31 Lalouschek W, Aull S, Serles W, Wolfsberger M, Deecke L, Pabinger-Fasching I, Mannhalter C. The relation between erythrocyte volume and folate levels is influenced by a common mutation in the methylenetetrahydrofolate reductase (MTHFR) gene (C677T).  J Investig Med. 2000;  48 14-20
• 32 Li Z, Sun L, Zhang H, Liao Y, Wang D, Zhao B, Zhu Z, Zhao J, Ma A, Han Y, Wang Y, Shi Y, Ye J, Hui R. Elevated plasma homocysteine was associated with hemorrhagic and ischemic stroke, but methylenetetrahydrofolate reductase gene C677T polymorphism was a risk factor for thrombotic stroke: a multicenter case-control study in China.  Stroke. 2003;  34 2085-2090
• 33 Lindpaintner K, Pfeffer M A, Kreutz R, Stampfer M J, Grodstein F, LaMotte F, Buring J, Hennekens C H. A prospective evaluation of an angiotensin-converting-enzyme gene polymorphism and the risk of ischemic heart disease.  N Engl J Med. 1995;  332 706-711
• 34 Montgomery H E, Clarkson P, Dollery C M, Prasad K, Losi M A, Hemingway H, Statters D, Jubb M, Girvain M, Varnava A, World M, Deanfield J, Talmud P, McEwan J R, McKenna W J, Humphries S. Association of angiotensin-converting enzyme gene I/D polymorphism with change in left ventricular mass in response to physical training.  Circul. 1997;  96 741-747
• 35 Pezzini A, Del Zotto E, Archetti S, Negrini R, Bani P, Albertini A, Grassi M, Assanelli D, Gasparotti R, Vignolo L A, Magoni M, Padovani A. Plasma homocysteine concentration, C677T MTHFR genotype, and 844ins68 bp CBS genotype in young adults with spontaneous cervical artery dissection and atherothrombotic stroke.  Stroke. 2002;  33 664-669
• 36 Rankinen T, Wolfarth B, Simoneau J A, Maier-Lenz D, Rauramaa R, Rivera M A, Boulay M R, Chagnon Y C, Perusse L, Keul J, Bouchard C. No association between the angiotensin-converting enzyme ID polymorphism and elite endurance athlete status.  J Appl Physiol. 2000;  88 1571-1575
• 37 Refsum H, Ueland P M, Nygard O, Vollset S E. Homocysteine and cardiovascular disease.  Annu Rev Med. 1998;  49 31-62
• 38 Rigat B, Hubert C, Alhenc-Gelas F, Cambien F, Corvol P, Soubrier F. An insertion/deletion polymorphism in the angiotensin I-converting enzyme gene account for half the variance of serum enzyme levels.  J Clin Invest. 1990;  86 1343-1346
• 39 Rigat B, Hubert C, Corvol P, Soubrier F. PCR detection of the insertion/deletion polymorphism of the human angiotensin-converting enzyme gene (DCP1) (dipeptidylcarboxypeptidase 1).  Nucl Acids Res. 1992;  20 1433
• 40 Russ A P, Maerz W, Ruzicka V, Stein U, Gross W. Rapid detection of the hypertension-associated Met235→Thr allele of the human angiotensinogen gene.  Hum Mol Genet. 1993;  2 609-610
• 41 Sacchi E, Tagliabue L, Duca F, Mannucci P M. High frequency of the C677T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene in Northern Italy.  Thromb Haemost. 1997;  78 963-964
• 42 Sarnak M J, Tighiouart H, Manjunath G, MacLeod B, Griffith J, Salem D, Levey A S. Anemia as a risk factor for cardiovascular disease in the Atherosclerosis Risk in Communities (ARIC) study.  J Am Coll Cardiol. 2002;  40 27-33
• 43 Sethi A A, Tybjaerg-Hansen A, Gronholdt M L, Steffensen R, Schnohr P, Nordestgaard B G. Angiotensinogen mutations and risk for ischemic heart disease, myocardial infarction, and ischemic cerebrovascular disease. Six case-control studies from the Copenhagen City Heart Study.  Ann Intern Med. 2001;  134 941-954
• 44 Shanmugam V, Sell K W, Saha B K. Mistyping ACE heterozygotes.  PCR Methods Appl. 1993;  3 120-121
• 45 Soriente L, Coppola A, Madonna P, Cerbone A M, Di Minno G, Orefice G, D'Angelo A. Homozygous C677T mutation of the 5,10 methylenetetrahydrofolate reductase gene and hyperhomocysteinemia in Italian patients with a history of early-onset ischemic stroke.  Stroke. 1998;  29 869-871
• 46 Tanis B C, Blom H J, Bloemenkamp D G, van den Bosch M A, Algra A, van der Graaf Y, Rosendaal F R. Folate, homocysteine levels, methylenetetrahydrofolate reductase (MTHFR) 677C→T variant, and the risk of myocardial infarction in young women: effect of female hormones on homocysteine levels.  J Thromb Haemost. 2004;  2 35-41
• 47 Ueland P M, Hustad S, Schneede J, Refsum H, Vollset S E. Biological and clinical implications of the MTHFR C677T polymorphism.  TRENDS Pharmacol Sci. 2001;  22 195-201
• 48 Wang W Y, Zee R Y, Morris B J. Association of angiotensin II type 1 receptor gene polymorphism with essential hypertension.  Clin Genet. 1997;  51 31-34
• 49 Williams A G, Rayson M P, Jubb M, World M, Woods D R, Hayward M, Martin J, Humphries S E, Montgomery H E. The ACE gene and muscle performance.  Nature. 2000;  403 614