Genetic variations of the endothelial nitric oxide synthase gene are related to increased levels of C-reactive protein and macrophage-colony stimulating-factor in patients with coronary artery disease

John P. Lekakis; Ignatios Ikonomidis; Maria Tsibida; Athanasios Protogerou; Aggeliki Papada; Aggeliki Papapanagiotou; Ioanna Revela; Christos M. Papamichael; Anastasios T. Kalofoutis; Dimitrios T. Kremastinos

doi:10.1160/TH06-05-0280

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2006; 96(04): 520-528
DOI: 10.1160/TH06-05-0280

Cardiovascular Biology and Cell Signalling

Schattauer GmbH

Genetic variations of the endothelial nitric oxide synthase gene are related to increased levels of C-reactive protein and macrophage-colony stimulating-factor in patients with coronary artery disease

John P. Lekakis

¹2^nd Department of Cardiology, Athens University, Athens, Greece

,

Ignatios Ikonomidis

¹2^nd Department of Cardiology, Athens University, Athens, Greece

,

Maria Tsibida

²Department of Biological Chemistry, Athens University, Athens, Greece

,

Athanasios Protogerou

³Department of Clinical Therapeutics, Alexandra University Hospital, Athens, Greece

,

Aggeliki Papada

²Department of Biological Chemistry, Athens University, Athens, Greece

,

Aggeliki Papapanagiotou

²Department of Biological Chemistry, Athens University, Athens, Greece

,

Ioanna Revela

³Department of Clinical Therapeutics, Alexandra University Hospital, Athens, Greece

,

Christos M. Papamichael

³Department of Clinical Therapeutics, Alexandra University Hospital, Athens, Greece

,

Anastasios T. Kalofoutis

²Department of Biological Chemistry, Athens University, Athens, Greece

,

Dimitrios T. Kremastinos

¹2^nd Department of Cardiology, Athens University, Athens, Greece

› Author Affiliations

Abstract

Summary

It was the objective of this study to investigate the relation between nitric oxide synthase (NOS3) gene polymorphisms, vascular inflammation, endothelial function, and atherosclerosis. We examined the effects of a variable nucleotide tandem repeats (VNTR ) in intron 4, G894T in exon 7 and T-786C at the promoter region of NOS3 on i) C-reactive protein (CRP) and macrophage-colony stimulating-factor (MCSF), and ii) augmentation index (AI) measured by pulse-wave analysis, flow-mediated dilation (FMD) of the brachial artery, intima-media thickness (IMT) of the carotid and femoral artery using ultrasonography and ankle-brachial index (ABI) in 122 patients with chronic coronary artery disease (CAD) who underwent coronary angiography. MCSF and CRP were increased in patients with T-786C (77/122) or VNTR (40/122) allele compared to those without (F=10.8, p=0.002 and F=3.8, p=0.04 for T-786C and F=3.65, p=0.04 and F=3.2 p=0.049 for VNTR), even after adjustment for traditional risk factors and medication. Patients with combination of VNTR and T-786C (31/122) had higher MCSF or CRP than patients with one or none of these alleles (p<0.05). Among patients with T-786C, those with MCSF>262 pg/ml or CRP>3.2 mg/l (n=33/77) had a higher femoral and carotid IMT and number of plaques in the peripheral arteries than those with lower values of these inflammatory indices (p<0.05). Patients with MCSF >262 pg/ml had also lower FMD and higher Gensini score than those with lower MCSF (p<0.05).The intron 4-VNTR and T-786C mutation of NOS3 gene enhance the inflammatory process in patients with chronic CAD.

Keywords

NO synthase gene polymorphisms - inflammation - atherosclerosis - vascular function

Full Text

References

References
1 Palmer RM, Ashton DS, Moncada S. Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature 1988; 333: 664-6.
2 Widlanski ME, Cokce N, Keany Jr. JF. et al. The clinical implications of endothelial dysfunction. J Am Coll Cardiol 2003; 42: 1149-60.
3 Peng HB, Rajavashisth TB, Libby P. et al. Nitric oxide inhibits macrophage-colony stimulating factor gene transcription in vascular endothelial cells. J Biol Chem 1995; 270: 17050-5.
4 Held TK, Mielke ME, Under M. et al. Kinetics and dose dependence of macrophage colony stimulating factor-induced proliferation and activation of murine mononuclear phagocytes in situ: differences between lungs, liver and spleen. J Interferon Cyt Res 1996; 16: 159-68.
5 Bataille R, Klein B. C-reactive protein levels as a direct indicator of interleukin 6 levels in humans in vivo . Arthritis Rheum 1992; 35: 982-4.
6 Hooper WC, Lally C, Austin H. et al. The relationship between polymorphisms in the endothelial cell nitric oxide synthase gene and the platelet GPIIIa gene with myocardial infarction and venous thromboembolism in African Americans. Chest 1999; 116: 880-6.
7 Poier O, Mao C, Mallet C. et al. .Polymorphisms of the endothelial nitric oxide synthase gene -No consistent association with myocardial infarction in the ECTIM study. Eur J Clin Invest 1999; 29: 284-90.
8 Wang LW, Wang J. Endothelial nitric oxide gene sequence variations and vascular disease. Mol Genet Metab 2000; 70: 241-51.
9 Kathiresan S, Otvos JD, Sullivan LM. et al. Common genetic variation at the endothelial nitric oxide gene synthase locus and relations to brachial artery vasodilator function in the community. Circulation 2005; 112: 1419-27.
10 Chrysohoou C, Panagiotakos DB, Pitsavos C. et al. Evidence for association between endothelial nitric oxide synthase gene polymorphism (G894T) and inflammatory markers: The ATTIKA study. Am Heart J 2004; 148: 733-8.
11 Lekakis JP, Papamichael CM, Vemmos CN. et al. Peripheral vascular endothelial dysfunction in patients with angina pectorisand normal coronary arteriograms. J Am Coll Cardiol 1998; 31: 541-6.
12 Lekakis JP, Papamichael CM, Cimponeriu AT. et al. Atherosclerotic changes of extracoronary arteries are associated with the extent of coronary atherosclerosis. Am J Cardiol 2000; 84: 949-52.
13 Papamichael CM, Lekakis JP, Stamatelopoulos KS. et al. Ankle-brachial index as a predictor of the extent of coronary atherosclerosis and cardiovascular events in patients with coronary artery disease. Am J Cardiol 2000; 86: 615-8.
14 Ikonomidis I, Lekakis I, Vamvakou G. et al. Cigarette smoking is associated with increased circulating proinflammatory and procoagulant markers in patients with chronic coronary artery disease: Effects of aspirin treatment. Am HeartJ 2005; 149: 832-9.
15 Pearson ES, Hartley HO. „Biometrika Tables for Statisticians Vol I”. 3rd Ed. Cambridge: Cambridge U. Press; 1970
16 Corretti MC, Anderson TJ, Benjamin EJ. et al. International Brachial Artery Reactivity Task Force. Guidelines for the ultrasound assessment of the endothelial-dependent flow-mediated vasodilation of the brachial artery: a report of the International Brachial Artery Reactivity Task Force. J Am Coll Cardiol 2002; 39: 257-65.
17 Lekakis JP, Papamichael CM, Barbaki P. et al. Comparison of low-density lipoprotein cholesterol lowering by pravastation to<100mg/dl versus >100mg/dl on brachial artery vasoreactivity in patients with severe hypercholesterolemia and previous atherosclerotic events or diabetes mellitus. Am J Cardiol 2002; 89: 857-60.
18 O’Rourke MF, Pauca A, Jiang XJ. Pulse wave analysis. Br J Clin Pharmacol 2001; 51: 507-22.
19 Miyahara K, Kawamoto T, Sase K. et al. Cloning and structural characterization of the human endothelial nitric-oxide-synthase gene. Eur J Biochem 1994; 223: 719-26.
20 Tsukada T, Yokoyama K, Arai T. et al. Evidence of association of the eNOS gene polymorphism with plasma NO metabolite levels in humans. Biochem Biophys Res Commun 1998; 245: 190-3.
21 Adachi T, Wang XL. Association of extracellular superoxide dismutase phenotype with the endothelial constitutive nitric oxide synthase polymorphism. FEBS Lett 1998; 433: 166-8.
22 Alexander RW, Dzau VJ. Vascular Biology. The past 50 years. Circulation 2000; 102: IV-112-IV-116.
23 Tesauro M, Thompson WC, Rogliani P. et al. Intracellular processing of endothelial nitric oxide synthase isoforms associated with differences in severity of cardiopulmonary diseases: cleavage of proteins with aspartate vs glutamate at positio 298. Proc Natl Acad Sci 2000; 97: 2832-5.
24 Leeson CP, Hingorani AD, Mullen MJ. et al. Glu298Asp endothelial nitric oxide synthase gene polymorphism interacts with enviromental and dietary factors to influence endothelial function. Circ Res 2002; 90: 1153-8.
25 Antoniades C, Tousoulis D, Vasiliadou C. et al. Genetic polymorphism on endothelial nitric oxide synthase affects endothelial activation and inflammatory response during the acute phase of myocardial infarction. J Am Coll Cardiol 2005; 46: 1101-9.
26 Miyamoto Y, Saito Y, Kajiyama N. et al. Endothelial nitric oxide synthase gene is positively associated with essential hypertension. Hypertension 1998; 32: 3-8.
27 Yoshimura M. A missense Glu298Asp variant in the endothelial nitric oxide synthase gene is associated with coronary spasm in the Japanese. Hum Genet 1998; 103: 65-9.
28 Cai H, Wilcken DE, Wang XL. The Glu298Asp (G894T) mutation at exon 7 of the endothelial nitric oxide synthase gene and coronary artery disease. J Mol Med 1999; 77: 511-4.
29 Lacolley P, Gautier S, Poitier O. et al. Nitric oxide synthase gene polymorphisms, blood pressure and aortic stiffness in normotensive and hypertensive subjects. J Hypertens 1998; 16: 31-5.
30 Nakayama M, Yasue H, Yoshimura M. et al. T-786C mutation in the 5’-flanking region of the endothelial nitric oxide synthase gene is associated with coronary spasm. Circulation 1999; 99: 2864-70.
31 Sim AS, Wang J, Wilcken D. et al. MspI polymorphism in the promoter of the human endothelial constitutive NO synthase gene in Australian Caucasian population. Mol Genet Metab 1998; 65: 62.
32 Rossi GP, Taddei S, Virdis A. et al. The T-786C and Glu298Asp polymorphisms of the endothelial nitric oxide gene affect the forearm blood flow responses of Caucasian hypertensive patients. J Am Coll Cardiol 2003; 41: 938-45.
33 Zhang C, Lopez-Ridaura R, Hunter DJ. et al. Common variants of the endothelial nitric oxide synthase gene and the risk of coronary heart disease among U.S. Diabetic men. Diabetes 2006; 55: 2140-7.
34 Yavuz D, Koc M, Toprak A. et al. Effects of ACE inhibition and AT1-receptor antagonism on endothelial function and insulin sensitivity in essential hypertensive patients. J Renin Angiotensin Aldosterone Syst 2003; 04: 197-203.
35 da Cunha V, Tham DM, Martin-McNulty B. et al. Enalapril attenuates angiotensin II-induced atherosclerosis and vascular inflammation. Atherosclerosis 2005; 178: 9-17.
36 Boger R, Baldus S. Clopidogrel improves systemic endothelial nitric oxide bioavailability in patients with coronary artery disease: evidence for antioxidant and antiinflammatory effects. Arterioscler Thromb Vasc Biol 2006; 26: 1648-52.
37 Karatzis E, Lekakis J, Papamichael C. et al. Rapid effect of pravastatin on endothelial function and lipid peroxidation in unstable angina. Int J Cardiol 2005; 101: 65-70.
38 Wu KK. Control of COX-2 and iNOS gene expressions by aspirin and salicylate. Thromb Res 2003; 110: 273-6.
39 Taubert D, Berkels R, Grosser N. et al. Aspirin induces nitric oxide release from vascular endothelium: a novel mechanism of action. Br J Pharmacol 2004; 143: 159-65.
40 Ridker PM. High-sensitivity C-reactive protein:potential adjunct for global risk assessment in the primary prevention of cardiovascular disease. Circulation 2001; 103: 1813-8.
41 Szmitko PE, Wang CH, Weisel RD. et al. New markers of inflammation and endothelial cell activation. Part I. Circulation 2003; 108: 1917-23.
42 Ikonomidis I, Lekakis J, Revela I. et al. Increased circulating C-reactive protein and macrophage -colony stimulating factor are complementary predictors of long-term outcome in patients with chronic coronary artery disease. Eur HeartJ 2005; 26: 1618-24.
43 Haraguchi K, Kubo M, Saito T. et al. Serum level of macrophage colony-stimulating factor and atherosclerosis in hemodialysis patients. Nephron Clin Pract 2006; 102: c14-20.
44 Matsui K, Yoshioka T, Murakami Y. et al. Serum concentrations of vascular endothelial growth factor and monocyte -colony stimulating factor are increased in peripheral artery disease. Circ J 2003; 67: 660-2.