Abstract
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
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Abbreviations used
- ESR
electron spin resonance
- ABTS
2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)
- HPE
human placenta extract
- RBC
red blood cell
Footnotes
Paper authored by participant of the international conference: 18th Meeting, European Association for Red Cell Research, Wrocław — Piechowice, Poland, May 12–15th, 2011. Publication cost was covered by the organizers of this meeting.
References
- 1.Halliwell B., Gutteridge J.M. Free radicals in biology and medicine. 2nd ed. Oxford and New York: Clarendon Press; 1989. pp. 23–30. [Google Scholar]
- 2.Jacob R.A., Burri B. Oxidative damage and defense. Am. J. Clin. Nutr. 1996;63:985–990. doi: 10.1093/ajcn/63.6.985. [DOI] [PubMed] [Google Scholar]
- 3.Solarska K., Lewińska A., Karowicz-Bilińska A., Bartosz G. The antioxidant properties of carnitine in vitro. Cell. Mol. Biol. Lett. 2010;15:90–97. doi: 10.2478/s11658-009-0036-y. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.May J.M., Qu Z., Xia L., Cobb C.E. Nitrite uptake and metabolism and oxidant stress in human erythrocytes. Am. J. Physiol. Cell. Physiol. 2000;279:1946–1954. doi: 10.1152/ajpcell.2000.279.6.C1946. [DOI] [PubMed] [Google Scholar]
- 5.Zavodnik I.B., Lapshina E.A., Rekawieckb K., Zavodnik L.B., Bartosz G., Bryszewska M. Membrane effects of nitrite-induced oxidation of human red blood cells. Biochim. Biophys. Acta. 1999;1421:306–316. doi: 10.1016/S0005-2736(99)00136-4. [DOI] [PubMed] [Google Scholar]
- 6.Shinde V., Dhalwal K., Paradkar A.R., Mahadik K.R., Kadam S.S. Evaluation of in-vitro antioxidant activity of human placental extract. Pharmacologyonline. 2006;3:172–179. [Google Scholar]
- 7.Rozanova S.L., Naumenko E.I., Rozanova E.D., Nardid O.A. Change of anioxidative properties of human placental extracts after freezing. Problems of Cryobiology. 2010;20:288–297. [Google Scholar]
- 8.Togashi S., Takahashi N., Iwama M., Watanabe S., Tamagawa K., Fukui T. Antioxidative collagen-derived peptides in human-placenta extract. Placenta. 2002;23:497–502. doi: 10.1053/plac.2002.0833. [DOI] [PubMed] [Google Scholar]
- 9.Shinde V. Effects of human placental extract on age related antioxidant enzyme status in D-galactose treated mice. Pharmacologyonline. 2007;1:252–261. [Google Scholar]
- 10.Park S.Y., Phark S., Lee M., Lim J.Y., Sul D. Anti-oxidative and antiinflammatory activities of placental extracts in benzo[a] pyrene-exposed rats. Placenta. 2010;31:873–879. doi: 10.1016/j.placenta.2010.07.010. [DOI] [PubMed] [Google Scholar]
- 11.Hong J.W., Lee W.J., Hahn S.B., Kim B.J., Lew D.H. The effect of human placenta extract in a wound healing model. Ann. Plast. Surg. 2010;65:96–100. doi: 10.1097/SAP.0b013e3181b0bb67. [DOI] [PubMed] [Google Scholar]
- 12.Re R., Pellegrini N., Proteggente A., Pannala A., Yang M., Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic. Biol. Med. 1999;26:1231–1237. doi: 10.1016/S0891-5849(98)00315-3. [DOI] [PubMed] [Google Scholar]
- 13.Harris D.A. Spectrophotometry and spectrofluorimetry. Washington: IRL Press; 1987. Spectrophotometric assays; pp. 49–90. [Google Scholar]
- 14.Arvint T., Cudd A., Shulz B., Nicolau C. Low-pH association of proteins with the membranes of intact red blood cells. Studies of the mechanism. Biochim. Biophys. Acta. 1989;981:51–61. doi: 10.1016/0005-2736(89)90081-3. [DOI] [PubMed] [Google Scholar]
- 15.Tsymbal L.V. Action of nonspecific factors on dynamic structure of the red blood cell cytoplasm. Problems of Cryobiology. 2000;3:8–15. [Google Scholar]
- 16.Henriquez C., Aliaga C., Lissi E. Kinetics profiles in the reaction of ABTS derived radicals with simple phenols and polyphenols. J. Chil. Chem. Soc. 2004;49:74–76. doi: 10.4067/S0717-97072004000100011. [DOI] [Google Scholar]
- 17.Nardid O.A. Effect of freezing on structural and functional properties of some hemoptoteins. Problems of Cryobiology. 1999;3:31–34. [Google Scholar]
- 18.Cao E., Chen Y., Cui Z., Forster P.R. Effect of freezing and thawing rates on denaturation of proteins in aqueous solutions. Biotechnol. Bioenerg. 2003;82:684–690. doi: 10.1002/bit.10612. [DOI] [PubMed] [Google Scholar]
- 19.Power G.G., Bragg S.L., Oshiro B.T., Dejam A., Hunter C.J., 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. 2007;103:1359–1365. doi: 10.1152/japplphysiol.00443.2007. [DOI] [PubMed] [Google Scholar]
- 20.Weichsel A., Kem M., Montfort W.R. Crystal structure of human thioredoxin revealing an unraveled helix and exposed S-nitrosation site. Protein Sci. 2010;19:1801–1806. doi: 10.1002/pro.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 21.Harwey, J.W. Erythrocyte Biochemistry in: Schalm’s Veterinary Hematology (Weiss, D., Wardrop, K.J. Eds.), 5th ed., 2010, 131–135.
