Koroner Kalp Hastalığında Endotelyal Nitrik Oksit Sentaz RS1799983 (Glu298asp) Varyasyonunun Metabolik Etkilerinin İncelenmesi

Yıl 2016, Cilt: 6 Sayı: 12, 21 - 31, 01.12.2016

Hülya Yılmaz Aydoğan , Serap İlikay Zehra Buğra Oğuz Öztürk

Öz

Amaç.
Koroner
kalp hastalığı (KKH) dünyada ölüm nedenlerinin başında gelen genetik ve çevresel

etkenlerin yol açtığı kompleks bir hastalıktır. KKH’na
yol açan ateroskleroz patogenezinde,
endotelyal

fonksiyon bozukluğu önemli bir anahtardır. Endotel hücreleri vasküler homeostaz ve nitrik oksit

üretimi gibi temel işlevlerini yerine getiremediğinde gelişen endotelyal disfonksiyon,
aterosklerotik

plak oluşumuna neden olur. Bu amaçla çalışmamızda eNOS (NOS3) geni rs1799983
(Glu298Asp) varyasyonunun

KKH riski açısından incelenmesi ve metabolik
etkilerinin gösterilmesi hedeflenmiştir.

Gereç
ve Yöntem. Çalışma gruplarımız 75 KKH ve 73 gönüllü sağlıklı kontrolden oluşturulmuştur.

NOS3 rs1799983 genotipleri PZR-RFLP
teknikleri kullanılarak belirlenmiştir.

Bulgular.
KKH hasta
ve kontrol gruplarında eNOS rs1799983 genotip ve
allel dağılımları benzerdir

(p>0.05). Çalışma gruplarında rs1799983 genotiplerinin
metabolik parametreler üzerine etkileri incelendiğinde,

kontrol grubunda eNOS nadir TT
(Asp/Asp) genotipi taşıyan bireylerde G (Glu) alleli
(GG+GT

genotipleri) taşıyanlara kıyasla vücut kitle indeksleri artmış bulundu. Ancak KKH hastalarında eNOS

rs1799983 varyasyonu metabolik ve
biyokimyasal parametrelerle ilişkili gözlenmemiştir (p>0.05).

Sonuç.
Bulgularımız eNOS rs1799983 varyasyonunun yüksek vücut kitle indeksi ile ilişkili olduğunu

düşündürmektedir.

Anahtar Kelimeler

eNOS, Gen, vücut kitle indeksi, metabolizma, Koroner kalp hastalığı

Metabolic Effects of the RS1799983 Variation (Glu298asp) of the Endothelial Nitric Oxide Synthase in Coronary Heart Disease

Öz

Objective.
Coronary heart disease (CHD) is a
complex disease caused by genetic and environmental

factors, being one of the major causes of death in the
world. Endothelial disfunction is important

key for atherosclerosis patogenesis leading to CHD.
Endothelial disfunction causes the formation of

atherosclerotic plaque when endothelial cells do not
perform ultimate functions such as vascular homeostasis

and nitric oxide (NO) production. In our study, we
purposed to investigate Endothelial NO

synthase (eNOS, NOS3) rs1799983 variation (Glu298Asp)
in CHD patients and determine its metabolic

effects on CHD development.

Material
and Methods. This study was carried out using
a sample of 75 CHD patients and 73

healthy controls. eNOS rs1799983 genotypes were
determined by polymerase chain reaction, restriction

fragment lenght polymorphism and agarose gel
electrophoresis.

Results. The eNOS rs1799983 genotype distributions were the
same between study groups (p>0.05).

When the effects of the rs1799983 genotypes were
examined on metabolic parameters in the study

groups, the body mass index was found to be increased
in the subjects with the TT (Asp/Asp) genotypes

as compared to G (Glu) allele (GG+GT genotypes).
However, eNOS rs1799983 was not associated

with metabolic and biochemical parameters in patients
with CHD (p>0.05).

Conclusion.
Our findings indicate that the
eNOS rs1799983 genotypes may be associated with

elevated body mass index.

Anahtar Kelimeler

eNOS, Gene, body mass index, Metabolism, Coronary heart disease

Kaynakça

• 1. Lahera V, Goicoechea M, de Vinuesa SG, Miana M, de las Heras N, Cachofeiro V, Luño J. Endothelial dysfunction, oxidative stress and inflammation in atherosclerosis: beneficial effects of statins.Curr Med Chem 2007;14(2):243-248. 2. Tritto I, Ambrosio G. The multi-faceted behavior of nitric oxide in vascular “inflammation”: catchy terminology or true phenomenon? Cardiovasc Res 2004;63:1-4. 3. Sitia S, Tomasoni L, Atzeni F, Ambrosio G, Cordiano C, Catapano A, Tramontana S, Perticone F, Naccarato P, Camici P, Picano E,Cortigiani L, Bevilacqua M, Milazzo L, Cusi D, Barlassina C, Sarzi-Puttini P, Turiel M. From endothelial dysfunction to atherosclerosis. Autoimmun Rev 2010;9(12):830-834. 4. Atochin DN, Huang PL. Endothelial nitric oxide synthase transgenic models of endothelia dysfunction. Pflugers Arch 2010;460:965- 974. 5. Marsden PA, Heng HH, Scherer SW, Stewart RJ, Hall AV, Shi XM, Tsui LC, Schappert KT. Structure and chromosomal localization of the human constitutive endothelial nitric oxide synthase gene. J Biol Chem 1993;268:17478-17488. 6. Chen PF, Tsai AL, Berka V, Wu KK. Endothelial Nitric-oxide Synthase Evidence for Bidomain Structure and Successful Reconstitution of Catalytic Activity from Two Separate Domains Generated by A Baculovirus Expression System. J Biol Chem 1996;271:14631-14635. 7. Venema RC, Sayegh HS, Kent JD, Harrison DG. Identification, Characterization and Comparison of the Calmodulin-binding Domains of the Endothelial and Inducible Nitric Oxide Synthases. J Biol Chem 1996;271:6435-6440. 8. Bode-Boger SM, Boger RH, Kienke S, Junker W, Frolich JC. Elevated L-arginine/dimethylarginine ratio contributes to enhanced systemic NO production by dietary L-arginine in hypercholesterolemic rabbits. Biochem Biophys Res Commun 1996;219:598-603. 9. Beckman JS, Koppenol WH. Nitric oxide, superoxide, and peroxynitrite: the good, the bad, and ugly. Am J Physiol 1996;271:1424-1437. 10. Arif Yalcin A, Faruk Akturk I, Celik O, Erturk M, Sabri Hancer V, Yalcin B, Isiksacan N, Uzun F, Ozbey Ozyilmaz S, Biyik I. Coronary artery ectasia is associated with the c.894G>T (Glu- 298Asp) polymorphism of the endothelial nitric oxide synthase gene. Tohoku J Exp Med 2014;232(2):137-44. 11. Luo JQ, Wen JG, Zhou HH, Chen XP, Zhang W. Endothelial nitric oxide synthase gene G894T polymorphism and myocardial infarction: a meta-analysis of 34 studies involving 21,068 subjects. PLoS One 2014;9(1):e87196. 12. Fedele F, Mancone M, Chilian WM, Severino P, Canali E, Logan S, De Marchis ML, Volterrani M, Palmirotta R, Guadagni F. Role of genetic polymorphisms of ion channels in the pathophysiology of coronary microvascular dysfunction and ischemic heart disease. Basic Res Cardiol 2013;108(6):387. 13. Heltianu C, Costache G, Gafencu A, Diaconu M, Bodeanu M, Cristea C, Azibi K, Poenaru L, Simionescu M. Relationship of eNOS gene variants to diseases that have in common an endothelial celldysfunction. J Cell Mol Med 2005;9(1):135-142. 14. Hingorani AD, Liang CF, Fatibene J, Lyon A, Monteith S, Parsons A, Haydock S, Hopper RV, Stephens NG, O’Shaughnessy KM, Brown MJ. A common variant of the endothelial nitric oxide synthase (Glu298-->Asp) is a major risk factor for coronary artery disease in the UK. Circulation 1999;100(14):1515-1520. 15. Colombo MG, Paradossi U, Andreassi MG, Botto N, Manfredi S, Masetti S, Biagini A, Clerico A. Endothelial nitric oxide synthase gene polymorphisms and risk of coronary artery disease. Clin Chem 2003;49(3):389-395. 16. Souza-Costa DC1, Belo VA, Silva PS, Sertorio JT, Metzger IF, Lanna CM, Machado MA, Tanus-Santos JE. Endothelial nitric oxide synthase genotype and ischemic heart disease: meta-analysis of 26 studies involving 23028 subjects. Circulation 2004;109:1359-1365. 17. Berdeli A, Sekuri C, Sirri Cam F, Ercan E, Sagcan A, Tengiz I, Eser E, Akin M. Association between the eNOS (Glu298Asp) and the RAS genes polymorphisms and premature coronary artery disease in a Turkish population. Clin Chim Acta 2005;351(1-2):87-94. 18. 18.Cam SF, Sekuri C, Tengiz I, Ercan E, Sagcan A, Akin M, Berdeli A. The G894T polymorphism on endothelial nitric oxide synthase gene is associated with premature coronary artery disease in a Turkish population. Thromb Res 2005;116(4):287-292 19. Granath B, Taylor RR, van Bockxmeer FM, Mamotte CD. Lack of evidence for association between endothelial nitric oxide synthase gene polymorphisms and coronary artery disease in the Australian Caucasian population. J Cardiovasc Risk 2001;8(4):235-241. 20. Caglayan AO, Kalay N, Saatci C, Yalcýn A, Akalýn H, Dundar M. Lack of association between the Glu298Asp polymorphism of endothelial nitric oxide synthaseand slow coronary flow in the Turkish population. Can J Cardiol. 2009;25(3):e69-72. 21. Guldiken B, Sipahi T, Guldiken S, Ustundag S, Budak M, Turgut N, Ozkan H. Glu298Asp polymorphism of the endothelial nitric oxide synthase gene in Turkish patients withischemic stroke. Mol Biol Rep 2009;36(6):1539-1543. 22. AfrasyapL, Ozturk G. NO level and endothelial NO synthase gene polymorphism (Glu298Asp) in the patients withcoronary artery disease from the Turkish population. Acta Biochim Biophys Sin (Shanghai). 2004;36(10):661-666. 23. Ragia G, Nikolaidis E, Tavridou A, Arvanitidis KI, Kanoni S, Dedoussis GV, Bougioukas G, Manolopoulos VG. Endothelial nitric oxide synthase gene polymorphisms 786T > C and 894G > T in coronary artery bypass graft surgery patients. Hum Genomics 2010;4(6):375-383. 24. Andrikopoulos GK, Grammatopoulos DK, Tzeis SE, Zervou SI, Richter DJ, Zairis MN, Gialafos EJ, Sakellariou DC, Foussas SG,Manolis AS, Stefanadis CI, Toutouzas PK, Hillhouse EW; GEMIG study investigators. Association of the 894G>T polymorphism in the endothelial nitric oxide synthase gene with risk of acute myocardial infarction. BMC Med Genet 2008;9:43. 25. Karvonen J, Kauma H, Kervinen K, Rantala M, Ikäheimo M, Päivänsalo M, Savolainen MJ, Kesäniemi YA. Endothelial nitric oxide synthase gene Glu298Asp polymorphism and blood pressure, leftventricular mass and carotid artery atherosclerosis in a population-based cohort. J Intern Med 2002;251(2):102-110. 26. 26.Aras O, Hanson NQ, Bakanay SM, Tsai MY, Gulec S. Endothelial nitric oxide gene polymorphism (Glu298Asp) is not associated with coronary artery disease in Turkish population. Thromb Haemost 2002;87(2):347-349. 27. Alp E, Menevse S, Tulmac M, Kan D, Yalcin R, Erkan AF, Cengel A. Lack of association between matrix metalloproteinase-9 and endothelial nitric oxide synthase gene polymorphisms and coronary artery disease in Turkish population. DNA Cell Biol 2009;28(7):343-350. 28. World Medical Association. World Medical Association Declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA 2013;310(20):2191- 2194. 29. Miyamoto Y, Saito Y, Kajiyama N, Yoshimura M, Shimasaki Y, Nakayama M, Kamitani S et al. Endothelial nitric oxide synthase gene is positively associated with essential hyper 30. Chilton RJ. Pathophysiology of Coronary Heart Disease: A Brief Review. JAOA 2004; (Supplement 7);104(9):S5-S8.18 31. Huang PL, Huang Z, Mashimo H, Bloch KD, Moskowitz MA, Bevan JA, Fishman MC. Hypertension in mice lacking the gene for endothelial nitric oxide synthase. Nature 1995;377:239-242. 32. Freedman JE, Sauter R, Battinelli EM, Ault K, Knowles C, Huang PL, Loscalzo J. Deficient platelet derived nitric oxide and enhanced hemostasis in mice lacking the NOSIII gene. Circ Res1999;84:1416-1421. 33. Lefer DJ, Jones SP, Girod WG, Baines A, Grisham MB, Cockrell AS, Huang PL, Scalia R. Leukocyte-endothelial cell interactions in nitric oxide synthase-deficient mice. Am J Physiol 1999;276:H1943–H1950. 34. Huang PL. Lessons learned from nitric oxide synthase knockout animals. Semin Perinatol 2000;24:87-90. 35. Huang Z, Huang PL, Ma J, Meng W, Ayata C, Fishman MC, Moskowitz MA. Enlarged infarcts in endothelial nitric oxide synthase knockout mice are attenuated by nitro-L-arginine. J Cereb Blood Flow Metab 1996;16:981-987. 36. Atochin DN, Wang A, Liu VW, Critchlow JD, Dantas AP, Looft-Wilson R, Murata T, Salomone S,Shin HK, Ayata C, Moskowitz MA, Michel T, Sessa WC, Huang PL. The phosphorylation state of eNOS modulates vascular reactivity and outcome of cerebral ischemia in vivo. J Clin Invest 2007;117:1961-1967. 37. Kuhlencordt PJ, Rosel E, Gerszten RE, Morales- Ruiz M, Dombkowski D, Atkinson WJ, Han F,Preffer F, Rosenzweig A, Sessa WC, Gimbrone MA Jr, Ertl G, Huang PL. Role of endothelial nitric oxide synthase in endothelial activation: insights from eNOS knockout endothelial cells. Am J Physiol Cell Physiol 2004;286:C1195–C1202. 38. Ohashi Y, Kawashima S, Hirata K, Yamashita T, Ishida T, Inoue N, Sakoda T, Kurihara H, Yazaki Y, Yokoyama M. Hypotension and reduced nitric oxide-elicited vasorelaxation in transgenic mice overexpressing endothelial nitric oxide synthase. J. Clin Invest 1998; 102(12):2061-2071. 39. Kawashima S, Yamashita T, Ozaki M, Ohashi Y, Azumi H, Inoue N, Hirata K, Hayashi Y, Itoh H, Yokoyama M. Endothelial no synthase overexpression inhibits lesion formation in mouse model of vascular remodeling. Arterioscler ThrombVasc Biol 2001;21:201-207. 40. Ozaki M, Kawashima S, Yamashita T, Hirase T, Namiki M, Inoue N, Hirata K, Yasui H, Sakurai H, Yoshida Y, Masada M, Yokoyama M. Overexpression of endothelial nitric oxide synthase accelerates atherosclerotic lesion formation in apoE-deficient mice. J Clin Invest. 2002;110(3):331-340. 41. Sansbury BE, Cumins TD, Tang Y, Hellman J, Holden CR, Harbeson MA, Chen Y, Patel RP, Spite MR, Bhatnager A, Hill BG. Overexpression of endothelial nitric oxide synthase prevents diet induced obesity and regulates adipocyte phenotype. Circ Res 2012;111(9):1176-1189. 42. Sessa WC. A new approach to weight loss: Just activate eNOS! Circ Res. 2012;111(9):1111-2. 43. Valerio A, Cardile A, Cozzi V, Bracale R, Tedesco L, Pisconti A, Palomba L, Cantoni O, Clementi E, Moncada S, Carruba MO, Nisoli E. Tnf-alpha downregulates enos expression and mitochondrial biogenesis in fat and muscle of obese rodents. J Clin Invest 2006;116:2791-2798. 44. Malhotra S, Poole J, Davis H, Dong Y, Pollock J, Snieder H, Treiber F. Effects of NOS3 Glu- 298Asp polymorphism on hemodynamic reactivity to stress: influences of ethnicity and obesity. Hypertension 2004;44: 866-871. 45. Souza-Costa DC , Belo VA , Silva P S , Metzger IF , Lanna CM , Machado MA, Tanus-Santos JE . eNOS haplotype associated with hypertension in obese children and adolescents. Int J Obes 2011;35:387-392. 46. Hingorani AD. Endothelial nitric oxide synthase polymorphisms and hypertension. Curr Hypertens Rep. 2003; 5:19-25.