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2006 | 53 | 2 |

Tytuł artykułu

Allelic polymorphism of endothelial NO-synthase gene and its functional manifestations

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
Investigation of the mechanisms of phenotypic realization of allelic polymorphism of the eNOS gene has shown that the level of eNOS mRNA and activity of this enzyme in platelets depends from genotype. We identified a T-786→ C polymorphism in the promoter region, a variable number of tandem repeats (4a/4b) in intron 4 and the G894→ T polymorphism in exon 7 of the eNOS gene in isolated human platelets. We measured eNOS mRNA in isolated platelets by reverse transcription-PCR and eNOS enzyme activity by fluorimetric detection system FCANOS-1 using diaminofluorescein diacetate (DAF-2A). It was shown that the level of eNOS mRNA is the lowest for the -786C/C promoter genotype. In exon 7 homozygotes (894G/G) the level of RNA is lower than in normal homozygotes (894G/G), but higher than in heterozygotes (894G/T). The eNOS activity in platelets is lower in carriers of the 786C/C promoter genotype than in normal homozygotes (2.1 × P=0.03), and lower comparing to heterozygotes (2.9 × P > 0.05). The eNOS activity accompanying the 894G/G variant of exon 7 is also lower than in normal homozygotes (P > 0.05). Regarding the polymorphism in intron 4 - the enzyme's activity is lower in carriers of the 4a/4a genotype comparing to normal homozygotes (1.7 × P > 0.05) and lower than in heterozygotes (1.9 × P > 0.05). These results allow one to conclude that the G-786→ C polymorphism of the eNOS gene promoter most significantly affects the gene expression and eNOS activity.

Wydawca

-

Rocznik

Tom

53

Numer

2

Opis fizyczny

p.299-302,fig.,ref.

Twórcy

  • Bogomoletz Institute of Physiology, Kiev, Ukraine
autor
autor

Bibliografia

  • Casas JP, Bautista LE, Humphries SE, Hingorani AD (2004) Endothelial nitric oxide synthase genotype and ischemic disease: meta-analysis of 26 studies involving 23028 subjects. Circulation 109: 1359–1365.
  • Dosenko VIe, Zahorii VIu, Moibenko OO, Parkhomenko OM (2002) Pathophysiologic aspects of endothelial Nosynthase genetic polymorphism. Fiziol Zh 48: 86–102 (in Ukrainian).
  • Dosenko VIe, Lutay YaM, Parkhomenko OM, Moibenko OO (2005) Endothelial NO-synthase allelic polymorphism frequency in patient with acute coronary syndrome in Ukrainian population. Tsytol Genet 2: 48–54 (in Ukrainian).
  • Golser R, Gorren ACF, Mayer B, Schmidt K (2003) Functional characterization of Glu298Asp mutant human endothelial nitric oxide synthase purified from a yeast expression system. Nitric Oxide 8: 7–14.
  • Hibi K, Ishigami T, Tamura K, Mizushima S, Nyui N, Fujita T, Fujita T, Ochiai H, Kosuge M, Watanabe Y, Yoshii Y, Kihara M, Kimura K, Ishii M, Umemura S (1998) Endothelial nitric oxide synthase gene polymorphism and acute myocardial infarction. Hypertension 32: 521–526.
  • Hingorani AD, Liang CF, Fatibene J, Lyon A, Monteith S, Parsons A, Haydock S, Hopper RV, Stephens NG, O’Shaughnessy KM, Brown MJ (1999) A common variant of the endothelial nitric oxide synthase (Glu298→ Asp) is a major risk factor for coronary artery disease in the UK. Circulation 100: 1515–1520.
  • Jayachandran M, Miller VM (2002) Ovariectomy upregulates expression of estrogen receptors, NOS, and HSPs in porcine platelets. Am J Physiol Heart Circ Physiol 283: H220–H226.
  • Nakayama M, Yasue H, Yoshimura M, Shimasaki Y, Kugiyama K, Ogawa H, Motoyama T, Saito Y, Ogawa Y, Miyamoto Y, Nakao K (1999) T-786→C Mutation in the 5'-flanking region of the endothelial nitric oxide synthase gene is associated with coronary spasm. Circulation 99: 2864–2870.
  • Oury C, Toth-Zsamboki E, Vermylen J, Hoylaerts MF (2002) P2X(1)-mediated activation of extracellular signal-regulated kinase 2 contributes to platelet secretion and aggregation induced by collagen. Blood 100: 2499–2505.
  • Øvstebø R, Haug KBF, Lande K, Kierulf P (2003) PCRbased calibration curves for studies of quantitative gene expression in human monocytes: development and evaluation. Clin Chem 49: 425–432.
  • Randriamboavonjy V, Schrader J, Busse R, Fleming I (2004) Insulin induces the release of vasodilator compounds from platelets by a nitric oxide–G kinase–VAMP-3–dependent pathway. J Exp Med 199: 347–356.
  • Song J, Yoon Y, Park KU, Park J, Hong YJ, Hong SH, Kim JQ (2003) Genotype-specific influence on nitric oxide synthase gene expression, protein concentration, and enzyme activity in cultured human endothelial cells. Clin Chem 49: 847–852.
  • Wang J, Dudley D, Wang XL (2002) Haplotype-specific effects on endothelial NO synthase promoter efficiency: modifiable by cigarette smoking. Arterioscler Thromb Vasc Biol 22: 1–4.
  • Wang XL, Sim AS, Wang MX, Murrell GA, Trudinger B, Wang J (2000) Genotype dependent and cigarette specific effects on endothelial nitric oxide synthase gene expression and enzyme activity. FEBS Lett 471: 45–50.
  • Yoon Y, Song J, Hong SH, Kim JQ (2000) Plasma nitric oxide concentrations and nitric oxide synthase gene polymorphisms in coronary artery disease. Clin Chem 46: 1626–1630.
  • Yoshimura M, Yasue H, Nakayama M, Shimasaki Y, Ogawa H, Kugiyama K, Saito Y, Miyamoto Y, Ogawa Y, Kaneshige T, Hiramatsu H, Yoshioka T, Kamitani S, Teraoka H, Nakao K (2000) Genetic risk factors for coronary artery spasm: significance of endothelial nitric oxide synthase gene T–786→C and missense Glu-298Asp variants. J Investig Med 48: 367–374.

Typ dokumentu

Bibliografia

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Identyfikator YADDA

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