Biological Trace Element Research

, Volume 189, Issue 2, pp 490–500 | Cite as

Protective Role of Selenium Against Hemolytic Anemia Is Mediated Through Redox Modulation

  • Rankaljeet Kaur
  • Preety Ghanghas
  • Pulkit Rastogi
  • Naveen KaushalEmail author


Selenium (Se), an essential trace element and potent nutritional antioxidant, exerts its biological effects through incorporation into selenoproteins like glutathione peroxidase (GPx). Modest decrement in the levels of GPx could be partly responsible for peroxidation of RBCs, which results into hemolytic anemia. Therefore, it is hypothesized that dietary Se, as selenoproteins (GPx), can maintain the homeostasis in RBCs and regulate the erythropoiesis by preventing oxidative stress-mediated hemolysis. Se-deficient (0.01 ppm), Se-adequate (0.1 ppm sodium selenite), and Se-supplemented (0.5 ppm sodium selenite) status were created in Balb/c mice by feeding yeast-based diets for 8 weeks and established by measuring Se levels in plasma and activities, expressions of Se-dependent selenoproteins. Fifty percent of mice from each differential Se group were treated with phenylhydrazine (PHZ, 20 mg/kg, i.p.) to induce hemolytic anemia. Results indicated that PHZ-treated Se-deficient animals demonstrated increased hemolysis, abnormal RBC morphology, increase in Heinz bodies and reticulocytes, and denaturation of hemoglobin to globin precipitates and methemoglobin. Se supplementation protected against these hemolytic changes and makes RBCs less fragile. These findings were consistent with dietary Se concentration-dependent changes in activity and expression of GPx indicating that ROS-mediated oxidative stress is integral to hemolysis. Protective effects of Se supplementation against increased levels of ROS, protein carbonyls, and peroxide damage to membrane lipids and enzymatic antioxidants validated these observations. In conclusion, dietary Se supplementation protected the RBCs against hemolysis by mitigating ROS-mediated oxidative stress.


Selenium Hemolytic anemia Oxidative stress Phenylhydrazine GPx 



Research reported in this publication was supported by the UGC-SAP (F.4-1/2015/DSA-1 (Sap-II)) and DST-FIST (SR/FST/LS1-645) programs sanctioned to the Department of Biophysics, Panjab University, Chandigarh (160014), India, by the University Grants Commission (UGC), Govt of India and the Department of Science and Technology (DST), Govt of India, respectively. The financial assistance to Panjab University by DST through DST-PURSE program is also duly acknowledged.

Compliance with Ethical Standards

All the experiments were performed in accordance with the guidelines of institutional ethical committee of Panjab University, Chandigarh.

Conflict Interest

The authors declare that they have no conflict of interest.

Supplementary material

12011_2018_1483_Fig7_ESM.png (1.1 mb)

(PNG 1092 kb)

12011_2018_1483_MOESM1_ESM.tif (135 kb)
High resolution image (TIF 135 kb)


  1. 1.
    Paqmantidis V, Bermano G, Villette S, Broom I, Arthur J, Hesketh J (2005) Effect of selenium depletion on glutathione peroxidase and selenoprotein W gene expression in the colon. FEBS Lett 579:792–796CrossRefGoogle Scholar
  2. 2.
    Navarro-Alarconm M, Cabrera-Vique C (2008) Selenium in food and the human body. A review. Sci Total Environ 400:115–141CrossRefGoogle Scholar
  3. 3.
    Hatfield DL, Berry MJ, Gladyshev VN (2001) Selenium: Its molecular biology and role in human health, pp 524–525Google Scholar
  4. 4.
    Rayman (2000) The importance of selenium to human health. Lancet 15:233–241CrossRefGoogle Scholar
  5. 5.
    Zhao J, Xing H, Liu C, Zhang Z, XU S (2016) Effect of selenium deficiency on nitric oxide and heat shock proteins in chicken erythrocytes. Biol Trace Elem Res 171:208–213CrossRefGoogle Scholar
  6. 6.
    Conrad M, Schweizer Y (2010) Unveiling the molecular mechanism behind selenium-related disease through knockout mouse studies. Antioxid Redox Signal 12:851–865CrossRefGoogle Scholar
  7. 7.
    Semba RD, Ricks MO, Ferrucci L, Xue QL, Guralnik JM, Fried LP (2009) Low serum selenium is associated with anemia among older adults in the United States. Eur J Clin Nutr 63:93–99CrossRefGoogle Scholar
  8. 8.
    Kupka R, Msamanga GI, Spiegelman D, Morris S, Mugusi F, Hunter DJ, Fawzi WW (2004) Selenium status is associated with accelerated HIV disease progression among HIV-1–infected pregnant women in Tanzania. J Nutr 134:2556–2560CrossRefGoogle Scholar
  9. 9.
    Ghaffari S (2008) Oxidative stress in the regulation of normal and neoplastic hematopoiesis. Antioxid Redox Signal 10(11):1923–1940CrossRefGoogle Scholar
  10. 10.
    Iuchi Y, Okada F, Onuma K, Onoda T, Asao H, Kobayashi M, Fujii J (2007) Elevated oxidative stress in erythrocytes due to an SOD1 deficiency causes anemia and triggers autoantibody production. Biochem J 402:219–227CrossRefGoogle Scholar
  11. 11.
    Fibach E, Rachmilewitz E (2008) The role of oxidative stress in hemolytic anemia. Curr Mol Med 8:609–619CrossRefGoogle Scholar
  12. 12.
    Fujii J, Kurahashi T, Konno T, Homma T, Luchi Y (2015) Oxidative stress as a potential causal factor for autoimmune hemolytic anemia and systemic lupus erythematosus. World J Nephrol 4:213–222CrossRefGoogle Scholar
  13. 13.
    Johnson RM, GJr G, Ravindranath Y, Ho YS (2005) Hemoglobin autoxidation and regulation of endogenous H2O2 levels in erythrocytes. Free Radic Biol Med 39:1407–1417CrossRefGoogle Scholar
  14. 14.
    Scott MD, Lubin BH, Zuo L, Kuypers FA (1991) Erythrocyte defense against hydrogen peroxide: preeminent importance of catalase. J Lab Clin Med 118:7–16Google Scholar
  15. 15.
    Klein EA (2004) Selenium: epidemiology and basic science. J Urol 171:S50–SS3Google Scholar
  16. 16.
    Kaushal N, Hegde S, Lumadue J, Paulson RF, Prabhu KS (2011) The regulation of erythropoiesis by selenium in mice. Antioxid Redox Signal 14:1403–1412CrossRefGoogle Scholar
  17. 17.
    Timothy M, Sheehan T, Gao M (1990) Simplified fluorometric assay of total selenium in plasma and urine. Clin Chem 36(/12):2124–2126Google Scholar
  18. 18.
    Paglia DE, Valentine WN (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 70:158–168Google Scholar
  19. 19.
    Holmgren A, Björnstedt M (1995) Thioredoxin and thioredoxin reductase. Methods Enzymol 252:199–208CrossRefGoogle Scholar
  20. 20.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with Folin-phenol reagent. J Biol Chem 193:265–275Google Scholar
  21. 21.
    Yeshoda KM (1942) Phenylhydrazine anaemia in rats. Curr Sci 11:360–363Google Scholar
  22. 22.
    Balasubramaniam P, Malathi A (1992) Comparative study of hemoglobin estimated by Drabkin’s and Sahli’s methods. J Postgrad Med 38:8–9Google Scholar
  23. 23.
    Sakata M, Yoshida A, Haga M (1982) Methemoglobin in blood as determined by double-wavelength spectrophotometry. Clin Chem 28:508–511Google Scholar
  24. 24.
    Sorensen S, Rubin E, Polster H, Mohandas N, Schrier S (1990) The role of membrane skeletal-associated alpha-globin in the pathophysiology of beta-thalassemia. Blood 75:1333–1336.1990Google Scholar
  25. 25.
    Best TM, Fiebig R, Corr DT, Brickson S, Brickson Ji L (1999) Free radical activity, antioxidant enzyme, and glutathione changes with muscle stretch injury in rabbits. J Appl Physiol 87:74–82CrossRefGoogle Scholar
  26. 26.
    Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG, Ahn BW, Shaltiel S, Stadtman ER (1990) Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol 186:464–478CrossRefGoogle Scholar
  27. 27.
    Wills ED (1966) Mechanisms of lipid peroxide formation in animal tissues. Biochem J 99(667–676):1966Google Scholar
  28. 28.
    Luck H (1971) Methods of enzymatic analysis. HU Bergmeyer Academic Press, New York, p 885Google Scholar
  29. 29.
    Kono Y (1978) Generation of superoxide radical during auto oxidation of hydroxylamine and an assay for superoxide dismutase. Arch Biochem Biophys 186:189–195CrossRefGoogle Scholar
  30. 30.
    Massey V, Williams CH Jr (1965) On the reaction mechanism of yeast glutathione reductase. J Biol Chem 240:4470–4480Google Scholar
  31. 31.
    Luangaram S, Kukoongviriyapan U, Pakdeechote P, Kukoongviriyapan V, Pannanqpetch P (2007) Protective effects of quercetin against phenylhydrazine-induced vascular dysfunction and oxidative stress in rats. Food Chem Toxicol 45:448–455CrossRefGoogle Scholar
  32. 32.
    Scott MD, Van R, Den Berg JJ, Repka T, Rouyer-Fessard P, Hebbel RP, Beuzard Y, Lubin BH (1993) Effect of excess alpha-hemoglobin chains on cellular and membrane oxidation in model beta-thalassemic erythrocytes. J Clin Investig 91:1706–1712CrossRefGoogle Scholar
  33. 33.
    Rotruck JT, Pope AL, Ganther HE, Hoekstra WG (1972) Prevention of oxidative damage to rat erythrocytes by dietary selenium. J Nutr 102:689–696CrossRefGoogle Scholar
  34. 34.
    Mcphail DB, Sibbald AM (1992) The role of free radicals in brassica-induced anaemia of sheep: an ESR spin trapping study. Free Radic Res Commun 16:277–284CrossRefGoogle Scholar
  35. 35.
    Sachdev SW, Sunde RA (2001) Selenium regulation of transcript abundance and translational efficiency of glutathione peroxidase-1 and -4 in rat liver. J Biochem 357:851–858Google Scholar
  36. 36.
    Rao L, Puschner B, Prolla TA (2001) Gene expression profiling of low selenium status in the mouse intestine: transcriptional activation of genes linked to DNA damage, cell cycle control and oxidative stress. J Nutr 131:3175–3181CrossRefGoogle Scholar
  37. 37.
    Harley JD, Mauer AM (1960) Studies on the formation of Heinz bodies. I. Methemoglobin Production and Oxyhemoglobin Destruction. Blood 16:1722–1735Google Scholar
  38. 38.
    Jacob HS (1970) Mechanism of Heinz body formation and attachment to red cell membrane. Semin Hematol 7:341–354Google Scholar
  39. 39.
    Advani R, Sorenson S, Shinar E, Lande W, Rachmilewitz E, Schrier SL (1992) Characterization and comparison of the red blood cell membrane damage in severe human alpha-and beta thalassemia. Blood 79:1058–1063Google Scholar
  40. 40.
    Scott MD, Eaton JW (1995) Thalassaemic erythrocytes: cellular suicide arising from iron and glutathione dependent oxidation reactions. Br J Haematol 91:811–819CrossRefGoogle Scholar
  41. 41.
    Shinar E, Rachmilewitz EA (1990) Oxidative denaturation of red blood cells in thalassemia. Semin Hematol 27:70–82Google Scholar
  42. 42.
    Urbankova L, Horky P, Skladanka J, Pribilova M, Smolikova V, Nevrkla P, Cernei N, Lackova Z, Hedbavny J, Ridoskova A, Adam V, Kopel P (2018) Antioxidant status of rats blood and liver affected by sodium selenite and selenium nanoparticles. PeerJ 6:e4862. CrossRefGoogle Scholar
  43. 43.
    Hamdy MM, Mosallam DS, Jamal AM, Rabie WA (2015) Selenium and vitamin E as antioxidants in chronic hemolytic anemia: are they deficient? A case-control study in a group of Egyptian children. J Adv Res 6:1071–1077CrossRefGoogle Scholar
  44. 44.
    Ines D, Sonia B, Riadh BM, Amel EG (2006) A comparative study of oxidant-antioxidant status in stable and active vitiligo patients. Arch Dermatol Res 298:147–152CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Rankaljeet Kaur
    • 1
  • Preety Ghanghas
    • 1
  • Pulkit Rastogi
    • 2
  • Naveen Kaushal
    • 1
    Email author
  1. 1.Department of BiophysicsPanjab UniversityChandigarhIndia
  2. 2.Department of HematologyPostgraduate Institute of Medical Education and Research, (PGIMER)ChandigarhIndia

Personalised recommendations