Cellular & Molecular Biology Letters

, Volume 17, Issue 2, pp 240–248 | Cite as

Protective effect of placenta extracts against nitrite-induced oxidative stress in human erythrocytes

  • Svitlana Rozanova
  • Yana Cherkashina
  • Svitlana Repina
  • Katherina Rozanova
  • Oleg Nardid
Research Article


Aqueous-saline human placenta extract (HPE) is known to possess antioxidant activity due to the high concentration of bioactive substances. This fact allows its application in clinical practice in order to treat oxidation-induced diseases. Extract antioxidant activity is mainly conditioned by proteins. Freezing of extracts has been shown to lead to their antioxidant activity increasing due to protein conformation changes.

Different biological models are widely used in order to evaluate efficacy of novel antioxidants and mechanisms of their action. One such model appears to be erythrocytes under nitrite-induced oxidative stress. Nitrite is known to be able to penetrate erythrocyte membrane and to oxidize hemoglobin. In order to investigate whether HPE is able to decrease nitrite-induced oxidative injuries and to evaluate the protein contribution to this process, spectrophotometric and electron spin resonance (ESR) assays were used.

Experimental data have revealed that antioxidant activity of extracts and of some of their fractions correlates with methemoglobin concentration lowering. Preliminary erythrocyte incubation with an extract fraction of 12 kDa allows preservation of the structural-dynamic cytosol state the closest to the control.

Key words

Human placenta extract Protein Antioxidant activity Oxidative stress Nitrite Erythrocyte Methemoglobin Cytosol Freezing Thawing 

Abbreviations used


electron spin resonance


2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)


human placenta extract


red blood cell


  1. 1.
    Halliwell, B. and Gutteridge, J.M. Free radicals in biology and medicine (Clarendon Press: Oxford and New York), 2nd ed., 1989, 23–30.Google Scholar
  2. 2.
    Jacob, R.A. and Burri, B. Oxidative damage and defense. Am. J. Clin. Nutr. 63 (1996) 985–990.Google Scholar
  3. 3.
    Solarska, K., Lewińska, A., Karowicz-Bilińska, A. and Bartosz, G. The antioxidant properties of carnitine in vitro. Cell. Mol. Biol. Lett. 15 (2010) 90–97.PubMedCrossRefGoogle Scholar
  4. 4.
    May, J.M., Qu, Z., Xia, L. and Cobb, C.E. Nitrite uptake and metabolism and oxidant stress in human erythrocytes. Am. J. Physiol. Cell. Physiol. 279 (2000) 1946–1954.Google Scholar
  5. 5.
    Zavodnik, I.B., Lapshina, E.A., Rekawieckb, K., Zavodnik, L.B., Bartosz, G. and Bryszewska, M. Membrane effects of nitrite-induced oxidation of human red blood cells. Biochim. Biophys. Acta 1421 (1999) 306–316.PubMedCrossRefGoogle Scholar
  6. 6.
    Shinde, V., Dhalwal, K., Paradkar, A.R., Mahadik, K.R. and Kadam, S.S. Evaluation of in-vitro antioxidant activity of human placental extract. Pharmacologyonline 3 (2006) 172–179.Google Scholar
  7. 7.
    Rozanova, S.L., Naumenko, E.I., Rozanova, E.D. and Nardid, O.A. Change of anioxidative properties of human placental extracts after freezing. Problems of Cryobiology 20 (2010) 288–297.Google Scholar
  8. 8.
    Togashi, S., Takahashi, N., Iwama, M., Watanabe, S., Tamagawa, K. and Fukui, T. Antioxidative collagen-derived peptides in human-placenta extract. Placenta 23 (2002) 497–502.PubMedCrossRefGoogle Scholar
  9. 9.
    Shinde, V. Effects of human placental extract on age related antioxidant enzyme status in D-galactose treated mice. Pharmacologyonline 1 (2007) 252–261.Google Scholar
  10. 10.
    Park, S.Y., Phark, S., Lee, M., Lim, J.Y. and Sul, D. Anti-oxidative and antiinflammatory activities of placental extracts in benzo[a] pyrene-exposed rats. Placenta 31 (2010) 873–879.PubMedCrossRefGoogle Scholar
  11. 11.
    Hong, J.W., Lee, W.J., Hahn, S.B., Kim, B.J. and Lew, D.H. The effect of human placenta extract in a wound healing model. Ann. Plast. Surg. 65 (2010) 96–100.PubMedCrossRefGoogle Scholar
  12. 12.
    Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M. and Rice-Evans, C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic. Biol. Med. 26 (1999) 1231–1237.PubMedCrossRefGoogle Scholar
  13. 13.
    Harris, D.A. Spectrophotometric assays in: Spectrophotometry and spectrofluorimetry (Washington: IRL Press), 1987, 49–90.Google Scholar
  14. 14.
    Arvint T., Cudd A., Shulz B. and Nicolau, C. Low-pH association of proteins with the membranes of intact red blood cells. Studies of the mechanism. Biochim. Biophys. Acta 981 (1989) 51–61.CrossRefGoogle Scholar
  15. 15.
    Tsymbal, L.V. Action of nonspecific factors on dynamic structure of the red blood cell cytoplasm. Problems of Cryobiology 3 (2000) 8–15.Google Scholar
  16. 16.
    Henriquez, C., Aliaga C. and Lissi E. Kinetics profiles in the reaction of ABTS derived radicals with simple phenols and polyphenols. J. Chil. Chem. Soc. 49 (2004) 74–76.CrossRefGoogle Scholar
  17. 17.
    Nardid, O.A. Effect of freezing on structural and functional properties of some hemoptoteins. Problems of Cryobiology 3 (1999) 31–34.Google Scholar
  18. 18.
    Cao, E., Chen, Y., Cui, Z. and Forster, P.R. Effect of freezing and thawing rates on denaturation of proteins in aqueous solutions. Biotechnol. Bioenerg. 82 (2003) 684–690.CrossRefGoogle Scholar
  19. 19.
    Power, G.G., Bragg, S.L., Oshiro, B.T., Dejam, A., Hunter, C.J. and Blood A.B. A novel method of measuring reduction of nitrite-induced methemoglobin applied to fetal and adult blood of humans and sheep. J. Appl. Physiol. 103 (2007) 1359–1365.PubMedCrossRefGoogle Scholar
  20. 20.
    Weichsel, A., Kem, M. and Montfort, W.R. Crystal structure of human thioredoxin revealing an unraveled helix and exposed S-nitrosation site. Protein Sci. 19 (2010) 1801–1806.PubMedCrossRefGoogle Scholar
  21. 21.
    Harwey, J.W. Erythrocyte Biochemistry in: Schalm’s Veterinary Hematology (Weiss, D., Wardrop, K.J. Eds.), 5th ed., 2010, 131–135.Google Scholar

Copyright information

© © Versita Warsaw and Springer-Verlag Wien 2012

Authors and Affiliations

  • Svitlana Rozanova
    • 1
  • Yana Cherkashina
    • 1
  • Svitlana Repina
    • 1
  • Katherina Rozanova
    • 1
  • Oleg Nardid
    • 1
  1. 1.Department of Cryobiophysics, Institute for Problems of Cryobiology and CryomedicineNAS of UkraineKharkivUkraine

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