Molecular and Cellular Biochemistry

, Volume 295, Issue 1–2, pp 45–52 | Cite as

Role of antioxidant systems, lipid peroxidation, and nitric oxide in postmenopausal osteoporosis

  • Salih OzgocmenEmail author
  • Huseyin Kaya
  • Ersin Fadillioglu
  • Rabia Aydogan
  • Zumrut Yilmaz


In this study we assessed activities of antioxidant enzymes, lipid peroxidation end-products, and nitric oxide (NO) levels in women with postmenopausal osteoporosis (PMO). Relationship between oxidative stress parameters and NO levels with bone mineral density (BMD) and clinical variables influencing bone mass and health related quality of life measures was also investigated in women with PMO.

Postmenopausal women (n = 87), aged 40–65, without previous diagnosis or treatment for osteoporosis and independent in daily living activities were included. BMD was measured at the lumbar spine and proximal femur using dual-X-ray absorptiometry (DXA). Erythrocyte catalase (CATe) enzyme activity, erythrocyte and plasma enzyme activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and lipid peroxidation end-product malondialdehyde (MDA) and nitrite/nitrate levels, by product of NO were studied. A total of 23 healthy non-porotic women were included as controls.

Women with PMO had significantly lower erythrocyte CATe enzyme activity and higher erythrocyte malondialdehyde (MDAe) and erythrocyte nitric oxide (NOe) levels in comparison to controls whereas erythrocyte SODe and GSH-Px enzyme activity was similar. In plasma, osteporotic women had significantly higher SOD enzyme activity and higher MDA levels whereas similar GSH-Px enzyme activity and NO levels compared to non-porotic controls. Significant correlation was found between erythrocyte SODe, CATe enzyme activity and NOe levels with proximal femur BMD. Some of the quality of life scores as pain, mental, and social functions correlated with antioxidant enzyme activities and NO levels.

Consequently, oxidative stress markers may be an important indicator for bone loss in postmenopausal women. Further researches assessing the oxidative stress markers and NO in bone tissue and changes with anti-osteoporotic drugs would be valuable to better understand the role of free radicals, antioxidants, and NO in the regulation of bone mass.


antioxidant enzyme malondialdehyde nitric oxide postmenopausal osteoporosis 



Reactive oxygen species


Nitric oxide synthase


Neuronal nitric oxide synthase


Inducible nitric oxide synthase


Endothelial nitric oxide synthase


Postmenopausal osteoporosis


Dual-X-ray absorptiometry


Quality of Life Questionnaire of the European Foundation for Osteoporosis-41 item


Type I cross-linked C-telopeptide






Erythrocyte catalase


Erythrocyte superoxide dismutase


Erythrocyte malondialdehyde


Erythrocyte glutathione peroxidase


Erythrocyte nitric oxide


Bone mineral density


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  1. 1.
    Raisz LG (2005) Pathogenesis of osteoporosis: concepts, conflicts and prospects J Clin Invest 115:3318–3325PubMedCrossRefGoogle Scholar
  2. 2.
    Garrett IR, Boyce BF, Oreffo ROC, Bonewald L, Poser J, Mundy GR (1990) Oxygen-derived free radicals stimulate osteoclastic bone resorption in rodent bone in vitro and in vivo J Clin Invest 85:632–639PubMedCrossRefGoogle Scholar
  3. 3.
    Bax BE, Alam ASMT, Banerji B, Bax CMR, Bevis PJR, Stevens CR, Moonga BS, Blake DR, Zaidi M (1992) Stimulation of osteoclastic bone resorption by hydrogen peroxide Biochem Biophys Res Commun 183:1153–1158PubMedCrossRefGoogle Scholar
  4. 4.
    Lean JM, Davies JT, Fuller K, Jagger CJ, Kirstein B, Partington GA, Urry ZL, Chambers TJ (2003) A crucial role for thiol antioxidants in estrogen-deficiency bone loss J Clin Invest 112:915–923PubMedCrossRefGoogle Scholar
  5. 5.
    Lean JM, Jagger CJ, Kirstein B, Fuller K, Chambers TJ (2005) Hydrogen peroxide is essential for estrogen-deficiency bone loss and osteoclasts formation Endocrinology 146:728–735PubMedCrossRefGoogle Scholar
  6. 6.
    Bai XC, Lu D, Bai J, Zheng H, Ke ZY, Li XM, Luo SQ (2004) Oxidative stress inhibits osteoblastic differentiation of bone cells by ERK and NF-κB Biochem Biophys Res Commun 314:197–207PubMedCrossRefGoogle Scholar
  7. 7.
    Park GB, Yoo CI, Kim HT, Kwon CH, Kim YK (2005) Role of mitogen-activated protein kinases in hydrogen peroxide-induced cell death in osteoblastic cells Toxicology 215:115–125PubMedCrossRefGoogle Scholar
  8. 8.
    Varanasi SS, Francis RM, Berger CEM, Papiha SS, Datta HK (1999) Mitochondrial DNA deletion associated oxidative stress and severe male osteoporosis Osteop Int 10:143–149CrossRefGoogle Scholar
  9. 9.
    Melhus H, Michaelsson K, Holmberg L, Wolk A, Ljunghall S (1999) Smoking, antioxidant vitamins and risk of hip fracture J Bone Miner Res 14:129–135PubMedCrossRefGoogle Scholar
  10. 10.
    Basu S, Michaelsson K, Olofsson H, Johansson S, Melhus H (2001) Association between oxidative stress and bone mineral density Biochem Biophys Res Commun 288:275–279PubMedCrossRefGoogle Scholar
  11. 11.
    Sontakke AN, Tare RS (2002) A duality in the roles of reactive oxygen species with respect to bone metabolism Clin Chim Acta 318:145–148PubMedCrossRefGoogle Scholar
  12. 12.
    Maggio D, Barabani B, Pierandrei M, Polidori MC, Catani M, Mecocci P, Senin U, Pacifici R, Cherubini A (2003) Marked decrease in plasma antioxidants in aged osteoporotic women. Results of a cross-sectional study JCEM 88:1523–1527PubMedGoogle Scholar
  13. 13.
    Yalin S, Bagis S, Polat G, Dogruer N, Aksit SC, Hatungil R, Erdogan C (2005) Is there a role of free oxygen radicals in primary male osteoporosis Clin Exp Rheumatol 23:689–692PubMedGoogle Scholar
  14. 14.
    Kigwell BA (2000) Nitric oxide-mediated metabolic regulation during exercise: effect of training in health and cardiovascular disease FASEB J 14:1685–1696CrossRefGoogle Scholar
  15. 15.
    Akyol O, Zoroglu SS, Armutcu F, Sahin S, Gurel A (2004) Nitric oxide as a physiopathological factor in neuropsychiatric disorders In vivo 18:377–390PubMedGoogle Scholar
  16. 16.
    van’t Hof RJ, Ralston SH (2001) Nitric oxide and bone Immunology 103:255–261PubMedCrossRefGoogle Scholar
  17. 17.
    Aguirre J, Buttery L, O’Shaughnessy M, Afzal F, Fernandez DMI, Hukkanen M, Huang P, MacIntyre I, Polak J (2001) Endothelial nitric oxide synthase gene-deficient mice demonstrate marked retardation in postnatal bone formation, reduced bone volume, and defects in osteoblast maturation and activity Am J Pathol 158:247–257PubMedGoogle Scholar
  18. 18.
    van’t Hof RJ, Armour KJ, Smith LM, Armour KE, Wei XQ, Liew FY, Ralston SH (2000) Requirement of the inducible nitric oxide synthase pathway for IL-1-induced osteoclastic bone resorption Proc Nat Acad Sci USA 97:7993–7998PubMedCrossRefGoogle Scholar
  19. 19.
    Caballero-Alias AM, Loveridge N, Lyon A, Das-Gupta V, Pitsillides A, Reeve J (2004) NOS isoforms in adult human osteocytes: multiple pathways of NO regulation? Calcif Tissue Int 75:78–84PubMedCrossRefGoogle Scholar
  20. 20.
    Leidig-Bruckner G, Genant HK, Minne HW, Storm T, Thamsborg G, Bruckner T, Sauer P, Schilling T, Soerensen OH, Ziegler R (1994) Comparison of a semiquantitative and a quantitative method for assessing vertebral fractures in osteoporosis Osteoporos Int 4:154–161PubMedCrossRefGoogle Scholar
  21. 21.
    Cerrahoglu L, Duruoz MT, Tikiz C, Olcenler S, Tulukoglu N, Susin A (2002) Postmenopozal kadinlarda diyetle kalsiyum alimi ile kemik mineral yogunlugu arasindaki iliski Ozteoporoz Dunyasindan 8:173–177Google Scholar
  22. 22.
    Kocyigit H, Gulseren S, Erol A, Hizli N, Memis A (2003) The reliability and validity of the Turkish version of Quality of Life Questionnaire of the European Foundation for Osteoporosis (QUALEFFO) Clin Rheumatol 22:18–23PubMedCrossRefGoogle Scholar
  23. 23.
    Aebi H (1974) Catalase. In: Bergmeyer HU (ed) Methods of Enzymatic Analysis. Academic Press, New York, pp. 673–677Google Scholar
  24. 24.
    Paglia DE, Valentine WN (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase J Lab Clin Med 70:158–170PubMedGoogle Scholar
  25. 25.
    Esterbauer H, Cheeseman KH (1990) Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. In: Packer L, Glazer AN (eds) Methods in Enzymology: V 186, Oxygen Radicals in Biological Systems. Academic Press, California, pp. 407–421Google Scholar
  26. 26.
    Sun Y, Oberley LW, Li Y (1988) A simple method for clinical assay of superoxide dismutase Clin Chem 34:497–500PubMedGoogle Scholar
  27. 27.
    Cortas NK, Wakid NW (1990) Determination of inorganic nitrate in serum and urine by a kinetic cadmium-reduction method Clin Chem 36:1440–1443PubMedGoogle Scholar
  28. 28.
    Hall TJ, Schaueblin M, Jeker H, Fuller K, Chambers TJ (1995) The role of reactive oxygen intermediates in osteoclastic bone resorption Biochem Biophys Res Commun 207:280–287PubMedCrossRefGoogle Scholar
  29. 29.
    Steinbeck MJ, Appel WH Jr, Verhoeven AJ, Karnovsky MJ (1994) NADPHoxidase expression and in situ production of superoxide by osteoclasts actively resorbing bone J Cell Biol 126:765–772PubMedCrossRefGoogle Scholar
  30. 30.
    Muthusami S, Ramachandran I, Muthusamy B, Vasudevan G, Prabhu V, Subramaniam V, Jagadeesan A, Narasimhan S (2005) Ovariectomy induces oxidative stress and impairs bone antioxidant system in adult rats Clin Chim Acta 360:81–86PubMedCrossRefGoogle Scholar
  31. 31.
    Jagger CJ, Lean JM, Davies JT, Chambers TJ (2005) Tumor necrosis factor-mediates osteopenia caused by depletion of antioxidants Endocrinology 146:113–118PubMedCrossRefGoogle Scholar
  32. 32.
    Kim HJ, Chang EJ, Kim HM, Lee SB, Kim HD, Kim GS, Kim HH (2006) Antioxidant α-lipoic acid inhibits osteoclast differentiation by reducing nuclear factor-κB DNA binding and prevents in vivo bone resorption induced by receptor activator of nuclear factor-κB ligand and tumor necrosis factor-α Free Rad Bio Med 40:1483–1493CrossRefGoogle Scholar
  33. 33.
    Wolf RL, Cauley JA, Pettinger M, Jackson R, Lacroix A, Leboff MS, Lewis CE, Nevitt MC, Simon JA, Stone KL, Wactawski-Wende J (2005) Lack of a relation between vitamin and mineral antioxidants and bone mineral density: results from the Women’s Health Initiative Am J Clin Nutr 82:581–588PubMedGoogle Scholar
  34. 34.
    Hao YJ, Tang Y, Chen FB, Pei FX (2005) Different doses of nitric oxide donor prevent osteoporosis in ovariectomized rats Clin Orthop Rel Res 435:226–231Google Scholar
  35. 35.
    van’t Hof RJ, Macphee J, Libouban H, Helfrich MH, Ralston SH (2004) Regulation of bone mass and bone turnover by neuronal nitric oxide synthase Endocrinology 145:5068–5074PubMedCrossRefGoogle Scholar
  36. 36.
    Caballero-Alias AM, Loveridge N, Lyon A, Das-Gupta V, Pitsillides A, Reeve J (2004) NOS isoforms in adult human osteocytes: multiple pathways of NO regulation? Calcif Tissue Int 75:78–84PubMedCrossRefGoogle Scholar
  37. 37.
    Jamal SA, Browner WS, Bauer DC, Cummings SR (1998) Intermittent use of nitrates increases bone mineral density: the study of osteoporotic fractures J Bone Miner Res 13:1755–1759PubMedCrossRefGoogle Scholar
  38. 38.
    Wimalawansa SJ (2000) Nitroglycerin therapy is as efficacious as standard estrogen replacement therapy (premarin) in prevention of oophorectomy-induced bone loss: a human pilot clinical study J Bone Miner Res 15:2240–2244PubMedCrossRefGoogle Scholar
  39. 39.
    Baecker N, Boese A, Schoenau E, Gerzer R, Heer M (2005) L-arginine, the natural precursor of NO, is not effective for preventing bone loss in postmenopausal women J Bone Miner Res 20:471–479PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2006

Authors and Affiliations

  • Salih Ozgocmen
    • 1
    Email author
  • Huseyin Kaya
    • 1
  • Ersin Fadillioglu
    • 2
  • Rabia Aydogan
    • 1
  • Zumrut Yilmaz
    • 3
  1. 1.Division of Rheumatology, Department of Physical Medicine and Rehabilitation, Faculty of MedicineFirat UniversityElazigTurkey
  2. 2.Department of Physiology, Faculty of MedicineHacettepe UniversityAnkaraTurkey
  3. 3.Department of Physiology, Faculty of MedicineInonu UniversityMalatyaTurkey

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