Elsevier

Biochemical Pharmacology

Volume 59, Issue 4, 15 February 2000, Pages 419-425
Biochemical Pharmacology

Hormones & Growth Factors
Protective effect of erythropoietin on the oxidative damage of erythrocyte membrane by hydroxyl radical

https://doi.org/10.1016/S0006-2952(99)00277-4Get rights and content

Abstract

Treatment of red blood cells with the copper (II) ascorbate system causes increased lipid peroxidation, increased membrane microviscosity, and phospholipid translocation with a concurrent decrease in cytosolic catalase and glutathione peroxidase activities. All these changes are prevented if the cells are treated with erythropoietin prior to the exposure to copper (II) ascorbate. The present investigation further indicates that the oxidative damage brought about by copper (II) ascorbate is due to generation of hydroxyl radical and that erythropoietin plays a unique role in protecting the membrane from oxidative damage.

Section snippets

Chemicals

Human recombinant erythropoietin, ascorbic acid, NADPH, BSA, bee venom PA2, phosphatidylserine, phosphatidylethanolamine, PC, catalase, reduced glutathione, oxidised glutathione, glutathione reductase, and DPH were procured from Sigma. Thiobarbituric acid and DMSO were purchased from E. Merck. Benzenesulfonic acid and Fast Blue BB salt were obtained from Aldrich. Other reagents used were of analytical grade.

Isolation of red blood cells

Goat blood was collected in acid–citrate dextrose solution. The packed RBC were isolated

Results

Copper (II) ascorbic acid has been found to increase lipid peroxidation of RBC membrane significantly, and administration of Ep prior to oxidative stress completely prevents lipid peroxidation (Fig. 1A). Follicle-stimulating hormone, another glycoprotein, exhibits no significant effect on copper (II) ascorbic acid-induced lipid peroxidation of RBC membrane (Fig. 1A). To understand and confirm whether the changes brought about by Ep were due to protection, the effect of the delayed addition of

Discussion

The use of copper (II) ascorbate as a free radical generating system is well established 22, 23, 24. In the present paper, free radicals were generated in vitro using the copper (II) ascorbate system. According to recent concepts [24], the metal-catalyzed autoxidation of ascorbate is accompanied by the reduction of the transition metal ions and the formation of hydrogen peroxide. Subsequently, the reduced metal ions react with hydrogen peroxide via the Fenton reaction: M(n−1)++H2O2→Mn++OH+OH·

Acknowledgements

The work was supported by the University Grants Commission (UGC), Government of India. Thanks are due to Dr. R. K. Banerjee for his constructive criticism and help in the preparation of the manuscript.

References (43)

  • C.F. Babbs et al.

    Detection and quantitation of hydroxyl radical using dimethyl sulfoxide as molecular probe

    Methods Enzymol

    (1990)
  • B. Halliwell et al.

    Oxygen free radicals and iron in relation to biology and medicineSome problems and concepts

    Arch Biochem Biophys

    (1986)
  • E.A. Lissi et al.

    Metabolic regulation in oxidative stressAn overview

  • V.B. Junqueira et al.

    Dose-dependent study of the effects of acute lindane administration on rat liver superoxide anion production, antioxidant enzyme activities and lipid peroxidation

    Toxicology

    (1986)
  • C.E. Cross et al.

    Antioxidant protectionA function of tracheobronchial and gastrointestinal mucus

    Lancet

    (1984)
  • P.H. Lai et al.

    Structural characterization of human erythropoietin

    J Biol Chem

    (1986)
  • D.A. Rowley et al.

    Superoxide-dependent and ascorbate-dependent formation of hydroxyl radicals in the presence of copper saltsA physiologically significant reaction

    Arch Biochem Biophys

    (1983)
  • B. Halliwell

    Antioxidant characterization

    Biochem Pharmacol

    (1995)
  • F. Stahlman

    Humoral regulation of erythropoiesis. XIV. A model for abnormal erythropoiesis in thalassemia

    Ann NY Acad Sci

    (1964)
  • H.P. Ward

    An in vitro assay of erythropoietin

    Proc Soc Exp Biol Med

    (1967)
  • S.B. Krantz et al.

    In vitro behavior of stem cells

    J Lab Clin Med

    (1968)
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