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Clinical Chemistry 51: 1159-1164, 2005. First published May 19, 2005; 10.1373/clinchem.2005.048850
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(Clinical Chemistry. 2005;51:1159-1164.)
© 2005 American Association for Clinical Chemistry, Inc.

Hemostasis and Thrombosis

Endothelial Nitric Oxide Synthase –786T>C, but Not 894G>T and 4a4b, Polymorphism Influences Plasma Homocysteine Concentrations in Persons with Normal Vitamin Status

Cinzia Fatini1,3,4,a, Francesco Sofi1,3,4, Anna Maria Gori1,3,4, Elena Sticchi1,3,4, Rossella Marcucci1,3,4, Meri Lenti1,3,4, Alessandro Casini2, Calogero Surrenti2, Rosanna Abbate1,3,4 and Gian Franco Gensini1,3,4

1 Department of Medical and Surgical Critical Care, 2 Department of Clinical Pathophysiology, Unit of Nutrition, and 4 Center for the Study at Molecular and Clinical Level of Chronic, Degenerative and Neoplastic Diseases to Develop Novel Therapies, University of Florence, Florence, Italy.
3 Thrombosis Centre, Azienda Ospedaliero-Universitaria Careggi, Florence, Italy.

aAddress correspondence to this author at: Department of Medical and Surgical Critical Care, Thrombosis Centre, University of Florence, Viale Morgagni 85, 50134 Florence, Italy. Fax 39-05-54279417; e-mail cinziafatini{at}hotmail.com.

Background: Nitric oxide (NO) plays a relevant role in various events during atherogenesis. In vitro data suggest that NO may modulate homocysteine (Hcy) concentrations. The aim of this study was to investigate the role of endothelial nitric oxide synthase (eNOS) –786T>C, 894G>T, and 4a4b polymorphisms in influencing Hcy concentrations.

Methods: Blood samples were obtained from 1287 unrelated persons. Plasma Hcy was measured by fluorescence polarization immunoassay, folate and vitamin B12 by RIA, vitamin B6 by HPLC, and eNOS and methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms by PCR with restriction fragment length polymorphism analysis.

Results: MTHFR 677C>T polymorphism significantly influenced Hcy concentrations after adjustment for all confounding variables (P <0.0001 for trend). Univariate analysis showed that the eNOS –786T>C polymorphism, but not 894G>T and 4a4b, was significantly associated with the risk of having Hcy in the third tertile [>13.4 µmol/L; odds ratio (OR) = 1.2; 95% confidence interval (CI), 1.02–1.5; P = 0.03]. After adjustment for all variables known to influence Hcy, the –786T>C polymorphism still influenced Hcy concentrations (OR = 1.9; 95% CI, 1.1–3.2; P = 0.01). By analyzing the influence of eNOS polymorphisms on plasma Hcy concentrations according to vitamin concentrations (folate, vitamin B6, and vitamin B12), age, and smoking habits, we found a significant association between the eNOS –786T>C polymorphism and Hcy in nonsmokers, in persons with normal vitamin status, and in persons <60 years.

Conclusion: The eNOS –786T>C polymorphism, but not 894G>T and 4a4b, influences plasma Hcy concentrations mildly but significantly and independently.




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