The Effects of Endurance Training and Vitamin D Supplementation on the Bone Resorption Markers in the Rats Exposed to Oxidative Damage Induced by Hydrogen Peroxide (H2O2)

Document Type : Research Paper


1 PhD of Exercise physiology, Shoushtar Branch, Islamic Azad University (I.A.U), Shoushtar, Iran

2 Professor of Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University (I.A.U), Tehran, Iran

3 Associate Professor of Neurosciences Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

4 Assistant Professor, Department of Physical Education & Sport Sciences , Masjed-Soleiman Branch, Islamic Azad University (I.A.U), Masjed-Soleiman, Iran


Introduction:studies have shown that the increase of active oxygen species is associated with damage to bone cells and the role of exercise and vitamin D on bone health has been proven;the aim of this study was to investigate the interactive effects of aerobic exercise with vitamin D supplements on bone resorption markers in rats exposed to oxidative damage induced by H2O2.
Methods: Sixty male adult male Wistar rats were randomly divided into 10 groups of six rats including:1)1 mmol/kg H2O2, 2)2 mmol/kg H2O2, 3)1mmol/kg H2O2+vitamin D,4) 1mmol/kg H2O2+aerobic training,5)1mmol/kg H2O2+Vitamin D+aerobic training,6)2 mmol/kg H2O2+Vitamin D,7)2mmol/kg H2O2+aerobic training,8)2mmol/kg H2O2+Vitamin D+aerobic training;9)Sham (D-methyl- Sulfoxide+normal saline); and(10)control and for 8 weeks were under the protocol of intervention.The levels of TRACP/5B and NTX were measured by ELISA method. Two-way and one-way ANOVA with Tukey’s post hoc test was used to analysis of data (p≤0.05).
Results: 1and 2 mmol/kg H2O2 had no significant effect on NTX and TRACP/5B levels(p≥0/05). Eight weeks of aerobic training increased of NTX levels in rats exposed to 2mmol/kg H2O2 (p≤0/05).Eight weeks of Vitamine D had no significant effect on NTX and TRACP/5B levels (p≥0/05).Also eight weeks of aerobic training along with Vitamin D had no interactive effects on increase of NTX and TRACP/5B levels in rats exposed to 1and 2mmol/kg H2O2(p≥0/05).
Conclusion: It seems that doses of 1 and 2mg/kg H2O2 have not significant effect on changes in bone resorption markers,but aerobic exercise had a significant effect on NTX increase in rats exposed to oxidative damage induced by 2mg/kg H2O2.


  1. 1. Domazetovic V, Marcucci G, Iantomasi T, Brandi ML, Vincenzini MT. Oxidative stress in bone remodeling: role of antioxidants. Clin Cases Miner Bone Metab. 2017; 14(2): 209–16.

    2. Liguori I, Russo G, Curcio F, Bulli G, Aran L, Della-Morte D, et al. Oxidative stress, aging, and diseases. Clin Interv Aging. 2018; 13: 757-72.

    3. Boyce BF, Li J, Xing L, Yao Z. Bone remodeling and the role of TRAF3 in osteoclastic bone resorption. Front Immunol. 2018; 9: 2263.

    4. Klein JA, Ackerman SL. Oxidative stress, cell cycle, and neurodegeneration. J Clin Invest. 2003; 111(6): 785–93.

    5. Sharma T, Islam N, Ahmad J, Akhtar N, Beg M. Correlation between bone mineral density and oxidative stress in postmenopausal women. Indian J Endocrinol Metab. 2015; 19(4): 491-7.

    6. Cervellati C, Bonaccorsi G, Cremonini E, Romani A, Fila E, Castaldini MC, et al. Oxidative stress and bone resorption interplay as a possible trigger for postmenopausal osteoporosis. Biomed Res Int. 2014; 2014: 569563.

    7. Jilka RL, Noble B, Weinstein RS. Osteocyte apoptosis. Bone. 2013; 54(2): 264–71.

    8. D’Oronzo S, Brown J, Coleman R. The role of biomarkers in the management of bone-homing malignancies. J Bone Oncol. 2017; 9: 1–9.

    9. Goudu AS, Naidub MD. Effect of fluoride on oxidative stress and biochemical markers of bone turnover on oxidative stress in postmenopausal women. Fluoride. 2013; 46(4): 208-11.‏

    1. Almeida M, Han L, Martin-Millan M, Plotkin LI, Stewart SA, Roberson PK, et al. Skeletal involution by age-associated oxidative stress and its acceleration by loss of sex steroids. J Biol Chem. 2007; 282(37): 27285–97.
    2. Bartell SM, Kim HN, Ambrogini E, Han L, Iyer S, Serra Ucer S, et al. FoxO proteins restrain osteoclastogenesis and bone resorption by attenuating H2O2 accumulation. Nat Commun. 2014; 5: 3773.
    3. Schwetz V, Trummer C, Pandis M, Grübler MR, Verheyen N, Gaksch M, et al. Effects of Vitamin D Supplementation on Bone Turnover Markers: A Randomized Controlled Trial. Nutrients. 2017; 9(5): E432.
    4. Tanzy ME, Camacho PM. Effect of vitamin D therapy on bone turnover markers in postmenopausal women with osteoporosis and osteopenia. Endocr Pract. 2011; 17(6): 873-9.
    5. Nahas-Neto J, Cangussu LM, Orsatti CL, Bueloni-Dias FN, Poloni PF, Schmitt EB, et al. Effect of isolated vitamin D supplementation on bone turnover markers in younger postmenopausal women: a randomized, double-blind, placebo-controlled trial. Osteoporos Int. 2018; 29(5): 1125-33.
    6. Hong AR, Kim SW. Effects of Resistance Exercise on Bone Health. Endocrinol Metab (Seoul). 2018; 33(4): 435–44.
    7. Shenoy S, Dhawan N, Sandhu JS. Effect of Exercise Program and Calcium Supplements on Low Bone Mass among Young Indian Women- A Comparative Study. Asian J SportsMed. 2012; 3(3): 193–9.
    8. Nebot E, Aparicio VA, Pietschmann P, Camiletti-Moirón D, Kapravelou G, Erben RG, et al. Effects of Hypertrophy Exercise in Bone Turnover Markers and Structure in Growing Male Rats. Int J Sports Med. 2017; 38(6): 418-25.
    9. Kim KH, Lee HB. Effects of circuit training interventions on bone metabolism markers and bone density of old women with osteopenia. J Exerc Rehabil. 2019; 15(2): 302–7.
    10. Pereira A, Costa AM, Palmeira-de-Oliveira A, Soares J, Monteiro M, Williams JHH. The effects of combined training on bone metabolic markers in postmenopausal women. Sci Sports. 2016; 31(3): 152-7.‏
    11. Romagnoli C, Marcucci G, Favilli F, Zonefrati R, Mavilia C, Galli G, et al. Role of GSH/GSSG redox couple in osteogenic activity and osteoclastogenic markers of human osteoblast-like SaOS-2 cells. FEBS J. 2013; 280(3): 867-79.
    12. Colares VLP, Lima SNL, Sousa NCF, Araújo MC, Pereira DMS, Mendes SJF, et al. Hydrogen peroxide-based products alter inflammatory and tissue damage-related proteins in the gingival crevicular fluid of healthy volunteers: a randomized trial. Sci Rep. 2019; 9(1): 3457.
    13. Zhou W, Liu Y, Shen J, Yu B, Bai J, Lin J et al. Melatonin Increases Bone Mass around the Prostheses of OVX Rats by Ameliorating Mitochondrial Oxidative Stress via the SIRT3/SOD2 Signaling Pathway. Oxid Med Cell Longev. 2019; 2019: 4019619.
    14. Soares DG, Gonçalves Basso F, Hebling J, de Souza Costa CA. Effect of hydrogen-peroxide-mediated oxidative stress on human dental pulp cells. J Dent. 2015; 43(6): 750-6.
    15. Fernandes MR, Barreto WDR Junior. Association between physical activity and vitamin D: A narrative literature review. Rev Assoc Med Bras (1992). 2017; 63(6): 550-6.‏
    16. Moosavi SJ, Habibian M, Farzanegi P. The effect of regular aerobic exercise on plasma levels of 25- hydroxy vitamin D and insulin resistance in hypertensive postmenopausal women with type 2 diabetes. RJMS. 2016; 22(141): 80-90. (Persian)
    17. Park H, Brannon PM, West AA, Yan J, Jiang X, Perry CA, et al. Maternal vitamin D biomarkers are associated with maternal and fetal bone turnover among pregnant women consuming controlled amounts of vitamin D, calcium, and phosphorus. Bone. 2017; 95: 183-91.‏
    18. Suzuki T, Nakamura Y, Kato H. Vitamin D and calcium addition during denosumab therapy over a period of four years significantly improves lumbar bone mineral density in Japanese osteoporosis patients. Nutrients. 2018; 10(3): E272.‏
    19. Tsuchiya Y, Sakuraba K, Ochi E. High force eccentric exercise enhances serum tartrate-resistant acid phosphatase-5b and osteocalcin. J Musculoskelet Neuronal Interact. 2014; 14(1): 50-7.‏
    20. Tong X, Chen X, Zhang S, Huang M1, Shen X1,3, Xu J, et al. The Effect of Exercise on the Prevention of Osteoporosis and Bone Angiogenesis. Biomed Res Int. 2019; 2019: 8171897.
    21. Nakamura Y, Kamimura M, Ikegami S, Mukaiyama K, Uchiyama S, Taguchi A, et al. Changes in serum vitamin D and PTH values using denosumab with or without bisphosphonate pre-treatment in osteoporotic patients: a short-term study. BMC Endocr Disord. 2015; 15: 81.
    22. Olmos JM, Hernández JL, Llorca J, Nan D, Valero C, González-Macías J. Effects of 25-hydroxyvitaminD3 therapy on bone turnover markers and PTH levels in postmenopausal osteoporotic women treated with alendronate. J Clin Endocrinol Metab. 2012; 97(12): 4491–7.
    23. Hedayati M, Rezaei N, Torkaman G, Movasseghe Sh, Bayat N. The Comparison of 6-Week Resistance Training and Pulsed Electromagnetic Field on Cortisol, and Anthropometric Parameters in Osteoporotic Postmenopausal Women. Iranian Journal of Endocrinology and Metabolism. 2012; 14(4): 380-91.
    24. Hassanzadeh H, Gozashti M, Dehkhoda M, Kazemi A. The Effect of Calcium and Vitamin D Consumption and Combined Training on Parathyroid Hormone and Alkaline Phosphatase of Postmenopausal Women. Medical journal of Mashhad University of medical sciences. 2012; 55(2): 96-101. (Persian)
    25. Matinhomaee H, Zobeiri M, Azarbayjani MA, Azizbeigi K. The effect of vitamin D supplementation during resistance training on the markers of systemic inflammation in untrained males. Scientific Journal of Kurdistan University of Medical Sciences. 2017; 21(6): 89-98. (Persian)