Journal of Bone and Mineral Metabolism

, Volume 32, Issue 5, pp 580–587 | Cite as

A susceptibility haplotype within the endothelial nitric oxide synthase gene influences bone mineral density in hypertensive women

  • Monica Singh
  • Puneetpal Singh
  • Surinder Singh
  • Pawan Kumar Juneja
  • Taranpal Kaur
Original Article


The influence of the coordinated effect of various single-nucleotide polymorphisms (SNPs) within the endothelial nitric oxide synthase (eNOS) gene on the risk of osteoporosis in hypertension has remained undetermined. Four pertinent SNPs of the eNOS gene, rs2070774, rs1799983, rs1800780 and rs3918181, were examined for the risk of osteoporosis in 313 hypertensive postmenopausal women in Northwest India. All the hypertensive women were verified with dual energy X-ray absorptiometry and categorized as 150 with osteoporosis and 163 without osteoporosis. The minor allele (T) of rs1799983 exerts a statistically significant risk for osteoporosis both in dominant [odds ratio (OR) 3.71, 95 % confidence interval (CI) 2.12–6.49, P < 0.001] and recessive mode (OR 5.75, 95 % CI 1.24–26.69, P = 0.036) after Bonferroni correction. Bone mineral density (BMD) values (corrected for the effects of risk variables) according to eNOS SNP genotypes revealed a significant association with rs1799983 at both the lumbar spine (P = 0.001) and femoral neck (P = 0.023). Risk association analyses revealed a susceptibility haplotype TTAG which influences the risk of osteoporosis (OR 2.02, 95 % CI 1.05–3.39, P = 0.042) in hypertension after adjusting for the effects of risk factors. Furthermore, this haplotype was significantly associated with BMD at the lumbar spine (P = 0.029) and femoral neck (P = 0.021) in a dose-dependent manner. The results suggest that possession of the TTAG haplotype of the eNOS gene may increase the risk of osteoporosis two-fold in hypertensive postmenopausal women in Northwest India.


eNOS gene Haplotype Bone mineral density Osteoporosis Hypertension 



The financial support of the DST project (SR/WOS-A/LS-387/2011) to MS is highly acknowledged.

Conflict of interest

All the authors have no conflict of interest.


  1. 1.
    Strazzullo P, Nunziata V, Cirillo M, Giannattasio R, Ferrara LA, Mattioli PL, Mancini M (1983) Abnormalities of calcium metabolism in essential hypertension. Clin Sci (Lond) 65:137–141Google Scholar
  2. 2.
    Hvarfner A, Bergstrom R, Morlin C, Wide L, Ljunghall S (1987) Relationship between calcium metabolic indices and blood pressure in patients with essential hypertension as compared with a healthy population. J Hypertens 5:451–456PubMedCrossRefGoogle Scholar
  3. 3.
    Wang TM, Hsu JF, Jee WS, Matthews JL (1993) Evidence for reduced cancellous bone mass in the spontaneously hypertensive rat. Bone Miner 20:251–264PubMedCrossRefGoogle Scholar
  4. 4.
    Ralston SH (1997) Science, medicine and the future: osteoporosis. BMJ 315:469–472PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Bucher HC, Cook RJ, Guyatt GH, Lang JD, Cook DJ, Hatala R, Hunt DL (1996) Effects of dietary calcium supplementation on blood pressure. A meta-analysis of randomized controlled trials. JAMA 275:1016–1022PubMedCrossRefGoogle Scholar
  6. 6.
    McCarron DA, Pingree PA, Rubin RJ, Gaucher SM, Molitch M, Krutzik S (1980) Enhanced parathyroid function in essential hypertension: a homeostatic response to a urinary calcium leak. Hypertension 2:162–168PubMedCrossRefGoogle Scholar
  7. 7.
    Cappuccio FP, Kalaitzidis R, Duneclift S, Eastwood JB (2000) Unravelling the links between calcium excretion, salt intake, hypertension, kidney stones and bone metabolism. J Nephrol 13:169–177PubMedGoogle Scholar
  8. 8.
    Tsuda K, Nishio I, Masuyama Y (2001) Bone mineral density in women with essential hypertension. Am J Hypertens 14:704–707PubMedCrossRefGoogle Scholar
  9. 9.
    MacGregor GA, Cappuccio FP (1993) The kidney and essential hypertension: a link to osteoporosis? J Hypertens 11:781–785PubMedCrossRefGoogle Scholar
  10. 10.
    Schoofs MW, van der Klift M, Hofman A, de Laet CE, Herings RM, Stijnen T, Pols HA, Stricker BH (2008) Thiazide diuretics and the risk for hip fracture. Ann Intern Med 139:476–482CrossRefGoogle Scholar
  11. 11.
    Schlienger RG, Kraenzlin ME, Jick SS, Meier CR (2004) Use of β-blockers and risk of fractures. JAMA 292:1326–1332PubMedCrossRefGoogle Scholar
  12. 12.
    von der Recke P, Hansen MA, Hassager C (1999) The association between low bone mass at the menopause and cardiovascular mortality. Am J Med 106:273–278PubMedCrossRefGoogle Scholar
  13. 13.
    Taylor BC, Schreiner PJ, Zmuda JM, Li J, Moffett SP, Beck TJ, Cummings SR, Lee JM, Walker K, Ensrud KE, SOF Research Group (2006) Association of endothelial nitric oxide Synthase genotypes with bone mineral density, bone loss, hip fracture and risk of fracture in older women: The SOF study. Bone 39:174–180Google Scholar
  14. 14.
    Cho K, Demissie S, Dupuis J, Cupples LA, Kathiresan S, Beck TJ, Karasik D, Kiel DP (2008) Polymorphisms in the endothelial nitric oxide synthase gene and bone density/ultrasound and geometry in humans. Bone 42:53–60PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Firat SC, Cetin Z, Samanci N, Aydin F, Balci N, Gungor F, Firat MZ, Luleci G, Karauzum SB (2009) Evaluation of eNOS gene polymorphisms in relation to BMD in postmenopausal women. Maturitas 63:352–356PubMedCrossRefGoogle Scholar
  16. 16.
    Liu SZ, Yan H, Hou WK, Xu P, Tian J, Tian LF, Zhu BF, Ma J, Lu SM (2009) Relationships between endothelial nitric oxide synthase gene polymorphisms and osteoporosis in postmenopausal women. J Zhejiang Uni Sci B 10:609–618CrossRefGoogle Scholar
  17. 17.
    Vallance P, Collier J, Moncada S (1989) Effects of endothelium-derived nitric oxide on peripheral arteriolar tone in man. Lancet 2:997–1000PubMedCrossRefGoogle Scholar
  18. 18.
    Hukkanen MV, Platts LA, Fernandez dMI, O’Shaughnessy M, MacIntyre I, Polak JM (1999) Developmental regulation of nitric oxide synthase expression in rat skeletal bone. J Bone Miner Res 14:868–877PubMedCrossRefGoogle Scholar
  19. 19.
    World Health Organization (WHO) (1993) Consensus development conference, diagnosis, prophylaxis and treatment of osteoporosis. Am J Med 94:646–650CrossRefGoogle Scholar
  20. 20.
    Srivastava K, Narang R, Sreenivas V, Das S, Das N (2008) Association of eNOS Glu298ASP gene polymorphism with essential hypertension in Asian Indians. Clin Chim Acta 387:80–83PubMedCrossRefGoogle Scholar
  21. 21.
    Excoffier L, Laval G, Schneider S (2007) Arlequin (version 3.0): an integrated software package for population genetics data analysis. Evol Bioinform Online 1:47–50PubMedCentralPubMedGoogle Scholar
  22. 22.
    Menashe I, Rosenberg PS, Chen BE (2008) PGA: power calculator for case-control genetic association analyses. BMC Genet 9:36PubMedCentralPubMedCrossRefGoogle Scholar
  23. 23.
    van’t Hof RJ, Ralston SH (2001) Nitric oxide and bone. Immunology 103:255–261PubMedCentralPubMedCrossRefGoogle Scholar
  24. 24.
    Huang PL, Huang Z, Mashimo H, Bloch KD, Moskowitz MA, Bevan JA, Fishman MC (1995) Hypertension in mice lacking the gene for endothelial nitric oxide synthase. Nature 377:239–242PubMedCrossRefGoogle Scholar
  25. 25.
    Alam I, Sun Q, Liu L, Koller DL, Fishburn T, Carr LG, Econs MJ, Foroud T, Turner CH (2006) Identification of a quantitative trait locus on rat chromosome 4 that is strongly linked to femoral neck structure and strength. Bone 39:93–99PubMedCrossRefGoogle Scholar
  26. 26.
    Li J, Cun Y, Tang WR, Wang Y, Li SN, Ouyang HR, Wu YR, Yu HJ, Xiao CJ (2011) Association of eNOS gene polymorphisms with essential hypertension in the Han population in southwestern China. Genet Mol Res 27:2202–2212CrossRefGoogle Scholar
  27. 27.
    Souza-Costa DC, Belo VA, Silva PS, Sertorio JT, Metzger IF, Lanna CM, Machado MA, Tanus-Santos JE (2011) eNOS haplotype associated with hypertension in obese children and adolescents. Int J Obes (Lond) 35:387–392CrossRefGoogle Scholar

Copyright information

© The Japanese Society for Bone and Mineral Research and Springer Japan 2013

Authors and Affiliations

  • Monica Singh
    • 1
  • Puneetpal Singh
    • 1
  • Surinder Singh
    • 2
  • Pawan Kumar Juneja
    • 2
  • Taranpal Kaur
    • 3
  1. 1.Molecular Genetics Laboratory, Department of Human GeneticsPunjabi UniversityPatialaIndia
  2. 2.Aggarwal Orthopedic HospitalLudhianaIndia
  3. 3.Amrit Sagar HospitalFerozepurIndia

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