Sport Sciences for Health

, Volume 15, Issue 3, pp 569–576 | Cite as

Administration of vitamin D and aerobic training: recovery of lung apoptosis markers in male rats exposed to hydrogen peroxide

  • Somayeh Ramezani
  • Maghsoud PeeriEmail author
  • Mohammad Ali Azarbayjani
  • Firouzeh Dehghan
Original Article



Apoptosis is one of the indications of programmed cell death (PCD) and is known as a physiological event in multicellular organisms. This study was designed to determine the effect of aerobic training alongside vitamin D supplementation on lung cell apoptosis in male rats exposed to hydrogen peroxide (H2O2).


48 male rats were assigned into six groups: H2O2, (H); H2O2 + D3, (HD); H2O2 + training, (HE); H2O2 + D3 + E, (HDE); dimethyl sulfoxide, (DMSO) and control intact. 1 mmol/kg of H2O2 was injected three times per week. The exercising rats performed the program on a rodent’s treadmill for 8 weeks, 5 days a week. The HD and HDE groups received a daily dose of 0.5 µg for 8 weeks. The lung tissue was exposed and stored at − 80. Then, the RT-PCR method was employed to examine the gene expressions of BAX, BCL2, Caspase-3 and Bcl-2/Bax ratios.


Results indicated that training, as well as a combination of training and vitamin D had a significant effect on BAX, BCL2 and Bel2/Bax ratio in case of H2O2 toxicity. The training and vitamin D groups both had no significant effect on Caspase-3 gene expression.


Based on the results of this research, it can be concluded that regular aerobic training alongside consumption of D3 might result in significant alteration of the genes involved in apoptosis caused by H2O2 presence in lung tissues.


Apoptosis Oxidative stress BCL2 BAX Caspase-3 Aerobic training Vitamin D 


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

For this type of study formal consent is not required.


  1. 1.
    Fedirko V, Bostick RM, Flanders WD, Long Q, Shaukat A, Rutherford RE, Daniel CR, Cohen V, Dash C (2009) Effects of vitamin D and calcium supplementation on markers of apoptosis in normal colon mucosa: a randomized, double-blind, placebo-controlled clinical trial. Cancer Prev Res 2(3):213–23Google Scholar
  2. 2.
    Savill J, Hogg N, Ren Y, Haslett C (1992) Thrombospondin cooperates with CD36 and the vitronectin receptor in macrophage recognition of neutrophils undergoing apoptosis. J Clin Investig 90:1513–1522PubMedGoogle Scholar
  3. 3.
    Lu Q, Harrington EO, Rounds S (2005) Apoptosis and lung injury. Keio J Med 54:184–189PubMedGoogle Scholar
  4. 4.
    Magi B, Bargagli E, Bini L, Rottoli P (2006) Proteome analysis of bronchoalveolar lavage in lung diseases. Proteomics 6:6354–6369PubMedGoogle Scholar
  5. 5.
    De Torre C, Ying SX, Munson PJ, Meduri GU, Suffredini AF (2006) Proteomic analysis of inflammatory biomarkers in bronchoalveolar lavage. Proteomics 6:3949–3957PubMedGoogle Scholar
  6. 6.
    Bargagli E, Olivieri C, Bennett D, Prasse A, Muller-Quernheim J, Rottoli P (2009) Oxidative stress in the pathogenesis of diffuse lung diseases: a review. Respir Med 103:1245–1256PubMedGoogle Scholar
  7. 7.
    Ye Q, Dalavanga Y, Poulakis N, Sixt SU, Guzman J, Costabel U (2008) Decreased expression of heme oxygenase-1 by alveolar macrophages in idiopathic pulmonary fibrosis. Eur Respir J 31(5):1030–6Google Scholar
  8. 8.
    Sato T, Takeno M, Honma K, Yamauchi H, Saito Y, Sasaki T, Morikubo H, Nagashima Y, Takagi S, Yamanaka K (2006) Heme oxygenase-1, a potential biomarker of chronic silicosis, attenuates silica-induced lung injury. Am J Respir Crit Care Med 174:906–914PubMedGoogle Scholar
  9. 9.
    Hatao H, Oh-ishi S, Itoh M, Leeuwenburgh C, Ohno H, Ookawara T, Kishi K, Yagyu H, Nakamura H, Matsuoka T (2006) Effects of acute exercise on lung antioxidant enzymes in young and old rats. Mech Ageing Dev 127:384–390PubMedGoogle Scholar
  10. 10.
    Aldini G, Yeum K-J, Niki E, Russell RM. Biomarkers for antioxidant defense and oxidative damage: principles and practical applications. John Wiley & Sons, HobokenGoogle Scholar
  11. 11.
    Gönenç A, Tokgöz D, Aslan S, Torun M (2005) Oxidative stress in relation to lipid profiles in different stages of breast cancer. Indian J Biochem Biophys 42(3):190–4Google Scholar
  12. 12.
    Franco R, Sánchez-Olea R, Reyes-Reyes EM, Panayiotidis MI (2009) Environmental toxicity, oxidative stress and apoptosis: menage a trois. Mutat Res 674:3–22Google Scholar
  13. 13.
    Uhal B (2008) The role of apoptosis in pulmonary fibrosis. Eur Respir Rev 17:138–144Google Scholar
  14. 14.
    Weinmann M, Jendrossek V, Handrick R, Güner D, Goecke B, Belka C (2004) Molecular ordering of hypoxia-induced apoptosis: critical involvement of the mitochondrial death pathway in a FADD/caspase-8 independent manner. Oncogene 23:3757PubMedGoogle Scholar
  15. 15.
    Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35:495–516PubMedPubMedCentralGoogle Scholar
  16. 16.
    Krüger K, Mooren FC (2014) Exercise-induced leukocyte apoptosis. Exerc Immunol Rev 20:117–134Google Scholar
  17. 17.
    Youle RJ, Strasser A (2008) The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 9:47PubMedGoogle Scholar
  18. 18.
    Wei MC, Zong W-X, Cheng EH-Y, Lindsten T, Panoutsakopoulou V, Ross AJ, Roth KA, MacGregor GR, Thompson CB, Korsmeyer SJ (2001) Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 292:727–730PubMedPubMedCentralGoogle Scholar
  19. 19.
    Polidoro L, Properzi G, Marampon F, Gravina G, Festuccia C, Di Cesare E, Scarsella L, Ciccarelli C, Zani B, Ferri C (2013) Vitamin D protects human endothelial cells from H2O2 oxidant injury through the Mek/Erk-Sirt1 axis activation. J Cardiovasc Transl Res 6:221–231PubMedGoogle Scholar
  20. 20.
    Tashkin DP, Celli B, Senn S, Burkhart D, Kesten S, Menjoge S, Decramer M (2008) A 4-year trial of tiotropium in chronic obstructive pulmonary disease. N Engl J Med 359:1543–1554PubMedGoogle Scholar
  21. 21.
    Kuepper T, Morrison A, Gieseler U, Schoeffl V (2009) Sport climbing with pre-existing cardio-pulmonary medical conditions. Int J Sports Med 30:395–402PubMedGoogle Scholar
  22. 22.
    Powers S, Nelson WB, Larson-Meyer E (2011) Antioxidant and vitamin D supplements for athletes: sense or nonsense? J Sports Sci 29(Suppl 1):S47–S55PubMedGoogle Scholar
  23. 23.
    Quadrilatero J, Alway SE, Dupont-Versteegden EE (2011) Skeletal muscle apoptotic response to physical activity: potential mechanisms for protection. Appl Physiol Nutr Metab 36:608–617PubMedGoogle Scholar
  24. 24.
    Garber CE, Blissmer B, Deschenes MR, Franklin BA, Lamonte MJ, Lee IM, Nieman DC, Swain DP (2011) American College of Sports Medicine position stand. Quantity and quality of exercise for developing and maintaining cardiorespiratory, musculoskeletal, and neuromotor fitness in apparently healthy adults: guidance for prescribing exercise. Med Sci Sports Exerc 43:1334–1359PubMedGoogle Scholar
  25. 25.
    Li SF, Liu HX, Zhang YB, Yan YC, Li YP (2010) The protective effects of alpha-ketoacids against oxidative stress on rat spermatozoa in vitro. Asian J Androl 12:247–256PubMedGoogle Scholar
  26. 26.
    Makino A, Skelton MM, Zou AP, Cowley AW Jr (2003) Increased renal medullary H2O2 leads to hypertension. Hypertension (Dallas, Tex: 1979) 42:25–30Google Scholar
  27. 27.
    Husain K, Hazelrigg SR (2002) Oxidative injury due to chronic nitric oxide synthase inhibition in rat: effect of regular exercise on the heart. Biochimica et biophysica Acta 1587:75–82PubMedGoogle Scholar
  28. 28.
    Halder SK, Sharan C, Al-Hendy A (2012) 1,25-dihydroxyvitamin D3 treatment shrinks uterine leiomyoma tumors in the Eker rat model. Biol Reprod 86:116PubMedPubMedCentralGoogle Scholar
  29. 29.
    Plant DR, Gregorevic P, Warmington SA, Williams DA, Lynch GS (2003) Endurance training adaptations modulate the redox-force relationship of rat isolated slow-twitch skeletal muscles. Clin Exp Pharmacol Physiol 30:77–81Google Scholar
  30. 30.
    Wong ML, Medrano JF (2005) Real-time PCR for mRNA quantitation. Biotechniques 39:75PubMedGoogle Scholar
  31. 31.
    Weiner P, Berar-Yanay N, Davidovich A, Magadle R, Weiner M (2000) Specific inspiratory muscle training in patients with mild asthma with high consumption of inhaled beta(2)-agonists. Chest 117:722–727Google Scholar
  32. 32.
    Schieber M, Chandel NS (2014) ROS function in redox signaling and oxidative stress. Curr Biol 24:R453–R462PubMedPubMedCentralGoogle Scholar
  33. 33.
    Wattenberg B, Lithgow T (2001) Targeting of C-terminal (tail)-anchored proteins: understanding how cytoplasmic activities are anchored to intracellular membranes. Traffic (Copenhagen Denmark); 2:66–71Google Scholar
  34. 34.
    McArdle WD, Katch FI, Katch VL. Exercise physiology: nutrition, energy, and human performance. Lippincott Williams & Wilkins, PhiladelphiaGoogle Scholar
  35. 35.
    Fernandes T, Magalhães FdC, ECd C, Oliveira EMd (2012) Aerobic exercise training inhibits skeletal muscular apoptotic signaling mediated by VEGF-VEGR2 in spontaneously hypertesive rats. Revista Brasileira de Medicina do Esporte 18:412–418Google Scholar
  36. 36.
    Phaneuf S, Leeuwenburgh C (2001) Apoptosis and exercise. Med Sci Sports Exerc 33:393–396PubMedGoogle Scholar
  37. 37.
    Siu PM, Bryner RW, Martyn JK, Alway SE (2004) Apoptotic adaptations from exercise training in skeletal and cardiac muscles. FASEB J 18:1150–1152PubMedGoogle Scholar
  38. 38.
    Allon N, Amir A, Manisterski E, Rabinovitz I, Dachir S, Kadar T (2009) Inhalation exposure to sulfur mustard in the guinea pig model: clinical, biochemical and histopathological characterization of respiratory injuries. Toxicol Appl Pharmacol 241:154–162PubMedGoogle Scholar
  39. 39.
    Islam T, Gauderman WJ, Berhane K, McConnell R, Avol E, Peters JM, Gilliland FD (2007) Relationship between air pollution, lung function and asthma in adolescents. Thorax 62:957–963PubMedPubMedCentralGoogle Scholar
  40. 40.
    Bagci E, Vodovotz Y, Billiar T, Ermentrout G, Bahar I (2006) Bistability in apoptosis: roles of Bax, Bcl-2, and mitochondrial permeability transition pores. Biophys J 90:1546–1559PubMedPubMedCentralGoogle Scholar
  41. 41.
    Skommer J, Wlodkowic D, Deptala A (2007) Larger than life: mitochondria and the Bcl-2 family. Leuk Res 31:277–286PubMedGoogle Scholar
  42. 42.
    Hildeman DA, Mitchell T, Aronow B, Wojciechowski S, Kappler J, Marrack P (2003) Control of Bcl-2 expression by reactive oxygen species. Proc Natl Acad Sci 100:15035–15040Google Scholar
  43. 43.
    Ahmad D, Bakairy AK, Katheri AM, Tamimi W (2015) MDM2 (RS769412) G> a polymorphism in cigarette smokers: a clue for the susceptibility to smoking and lung cancer risk. Asian Pac J Cancer Prev 16:4057–4060PubMedGoogle Scholar
  44. 44.
    Packer N, Pervaiz N, Hoffman-Goetz L (2010) Does exercise protect from cognitive decline by altering brain cytokine and apoptotic protein levels? A systematic review of the literature. Exerc Immunol Rev 16:138–162Google Scholar
  45. 45.
    Fisher G, Schwartz DD, Quindry JC, Barberio MD, Foster EB, Jones KW, Pascoe DD (2010) Lymphocyte enzymatic antioxidant responses to oxidative stress following high-intensity interval exercise. Am J Physiol-Heart Circul Physiol 110:730–737Google Scholar
  46. 46.
    Pope CA, Burnett RT, Thun MJ, Calle EE, Krewski D, Ito K, Thurston GD (2002) Lung cancer, cardiopulmonary mortality, and long-term exposure to fine particulate air pollution. JAMA 287:1132–1141Google Scholar
  47. 47.
    Hoffman-Goetz L, Pervaiz N, Guan J (2009) Voluntary exercise training in mice increases the expression of antioxidant enzymes and decreases the expression of TNF-α in intestinal lymphocytes. Brain Behav Immun 23:498–506Google Scholar
  48. 48.
    Quadrilatero J, Bombardier E, Norris SM, Talanian JL, Palmer MS, Logan HM, Tupling AR, Heigenhauser GJ, Spriet LL (2009) Prolonged moderate-intensity aerobic exercise does not alter apoptotic signaling and DNA fragmentation in human skeletal muscle. Am J Physiol Endocrinol Metab 298:E534–E547PubMedGoogle Scholar
  49. 49.
    Adams JM, Cory S (1998) The Bcl-2 protein family: arbiters of cell survival. Science 281:1322–1326PubMedGoogle Scholar
  50. 50.
    Salakou S, Kardamakis D, Tsamandas AC, Zolota V, Apostolakis E, Tzelepi V, Papathanasopoulos P, Bonikos DS, Papapetropoulos T, Petsas T (2007) Increased Bax/Bcl-2 ratio up-regulates caspase-3 and increases apoptosis in the thymus of patients with myasthenia gravis. In vivo 21:123–132PubMedGoogle Scholar
  51. 51.
    Jacobson MD, Raff MC (1995) Programmed cell death and Bcl-2 protection in very low oxygen. Nature 374:814PubMedGoogle Scholar
  52. 52.
    Zadeh-Vakili A, Tehrani FR, Daneshpour MS, Zarkesh M, Saadat N, Azizi F (2013) Genetic polymorphism of vitamin D receptor gene affects the phenotype of PCOS. Gene 515:193–196Google Scholar
  53. 53.
    Zhu S, Li M, Figueroa BE, Liu A, Stavrovskaya IG, Pasinelli P, Beal MF, Brown RH, Kristal BS, Ferrante RJ (2004) Prophylactic creatine administration mediates neuroprotection in cerebral ischemia in mice. J Neurosci 24:5909–5912PubMedPubMedCentralGoogle Scholar
  54. 54.
    Deminice R, Rosa FT, Franco GS, Jordao AA, de Freitas EC (2013) Effects of creatine supplementation on oxidative stress and inflammatory markers after repeated-sprint exercise in humans. Nutrition 29:1127–1132PubMedGoogle Scholar
  55. 55.
    Marfe G, Manzi V, Tafani M, Pucci B, Gambacurta A, Russo M, Sinibaldi Salimei P (2012) The modulation of sirtuins and apoptotic proteins in rats after exhaustive exercise. Open J Mol Integr Physiol 2:65–74Google Scholar
  56. 56.
    Umegaki K, Higuchi M, Inoue K, Esashi T (1998) Influence of one bout of intensive running on lymphocyte micronucleus frequencies in endurance-trained and untrained men. Int J Sports Med 19:581–585PubMedGoogle Scholar
  57. 57.
    Thompson D, Williams C, McGregor SJ, Nicholas CW, McArdle F, Jackson MJ, Powell JR (2001) Prolonged vitamin C supplementation and recovery from demanding exercise. Int J Sport Nutr Exerc Metab 11:466–481PubMedGoogle Scholar
  58. 58.
    Gomez-Cabrera M-C, Domenech E, Romagnoli M, Arduini A, Borras C, Pallardo FV, Sastre J, Vina J (2008) Oral administration of vitamin C decreases muscle mitochondrial biogenesis and hampers training-induced adaptations in endurance performance. Am J Clin Nutr 87:142–149PubMedGoogle Scholar
  59. 59.
    Wadley GD, McConell GK (2010) High-dose antioxidant vitamin C supplementation does not prevent acute exercise-induced increases in markers of skeletal muscle mitochondrial biogenesis in rats. J Appl Physiol 108:1719–1726PubMedGoogle Scholar
  60. 60.
    Simon-Schnass I, Pabst H (1988) Influence of vitamin E on physical performance. Int J Vitam Nutr Res 58:49–54PubMedGoogle Scholar
  61. 61.
    Gomes E, Allgrove J, Florida-James G, Stone V (2011) Effect of vitamin supplementation on lung injury and running performance in a hot, humid, and ozone-polluted environment. Scand J Med Sci Sports 21:e452–e460PubMedGoogle Scholar
  62. 62.
    Sumida S, Tanaka K, Kitao H, Nakadomo F (1989) Exercise-induced lipid peroxidation and leakage of enzymes before and after vitamin E supplementation. Int J Biochem 21:835–838PubMedGoogle Scholar
  63. 63.
    Buzina R, Suboticanec K (1985) Vitamin C and physical working capacity. Int J Vitam Nutr Res Suppl 27:157–166PubMedGoogle Scholar
  64. 64.
    Close GL, Ashton T, Cable T, Doran D, Holloway C, McArdle F, MacLaren DP (2006) Ascorbic acid supplementation does not attenuate post-exercise muscle soreness following muscle-damaging exercise but may delay the recovery process. Br J Nutr 95:976–981PubMedGoogle Scholar

Copyright information

© Springer-Verlag Italia S.r.l., part of Springer Nature 2019

Authors and Affiliations

  • Somayeh Ramezani
    • 1
  • Maghsoud Peeri
    • 1
    Email author
  • Mohammad Ali Azarbayjani
    • 1
  • Firouzeh Dehghan
    • 1
  1. 1.Department of Exercise Physiology, Faculty of Physical Education and Sport Sciences, Central Tehran BranchIslamic Azad UniversityTehranIran

Personalised recommendations