Hormones & Growth FactorsProtective effect of erythropoietin on the oxidative damage of erythrocyte membrane by hydroxyl radical
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:
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)
- et al.
On the mechanism of erythropoietin-induced differentiation. IX. Induced synthesis of 9S ribonucleic acid of hemoglobin
J Biol Chem
(1971) - et al.
The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes
Arch Biochem
(1963) - et al.
A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase
J Biol Chem
(1952) - et al.
Spectrin as a stabilizer of the phospholipid asymmetry in the human erythrocyte membrane
Biochim Biophys Acta
(1978) - et al.
Improved procedure for the extraction of lipids from human erythrocytes
J Lipid Res
(1965) - et al.
The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid
J Biol Chem
(1960) - et al.
Effects of modulating erythrocyte membrane cholesterol on Rh0(D) antigen expression
Biochem Biophys Res Commun
(1980) - et al.
Dynamics of the hydrocarbon layer in liposomes of lecithin and sphingomyelin containing dicetylphosphate
J Biol Chem
(1974) - et al.
Leshmania donovaniRole of microviscosity of macrophage membrane in the process of parasite attachment and internalization
Exp Parasitol
(1988) - et al.
Microsomal lipid peroxidation
Methods Enzymol
(1978)
Detection and quantitation of hydroxyl radical using dimethyl sulfoxide as molecular probe
Methods Enzymol
Oxygen free radicals and iron in relation to biology and medicineSome problems and concepts
Arch Biochem Biophys
Metabolic regulation in oxidative stressAn overview
Dose-dependent study of the effects of acute lindane administration on rat liver superoxide anion production, antioxidant enzyme activities and lipid peroxidation
Toxicology
Antioxidant protectionA function of tracheobronchial and gastrointestinal mucus
Lancet
Structural characterization of human erythropoietin
J Biol Chem
Superoxide-dependent and ascorbate-dependent formation of hydroxyl radicals in the presence of copper saltsA physiologically significant reaction
Arch Biochem Biophys
Antioxidant characterization
Biochem Pharmacol
Humoral regulation of erythropoiesis. XIV. A model for abnormal erythropoiesis in thalassemia
Ann NY Acad Sci
An in vitro assay of erythropoietin
Proc Soc Exp Biol Med
In vitro behavior of stem cells
J Lab Clin Med
Cited by (143)
Oleic acid prevents erythrocyte death by preserving haemoglobin and erythrocyte membrane proteins
2023, Free Radical Biology and MedicineDrug delivery platforms for neonatal brain injury
2021, Journal of Controlled ReleaseAttenuation of arsenic induced high fat diet exacerbated oxidative stress mediated hepatic and cardiac injuries in male Wistar rats by piperine involved antioxidative mechanisms
2020, Food and Chemical ToxicologyCitation Excerpt :The obtained enzyme activity was expressed as μmoles H2O2 consumed/min/mg of protein. The activity of glutathione peroxidase (GPx) was measured from the cytosolic fraction of hepatic and cardiac tissues using the method of Paglia and Valentine (1967) with some modification as adopted by Chattopadhyay et al. (2000). The enzymatic activity was expressed as U/mg of protein.
Insights into the ameliorative effect of oleic acid in rejuvenating phenylhydrazine induced oxidative stress mediated morpho-functionally dismantled erythrocytes
2020, Toxicology ReportsCitation Excerpt :Despite morphometric differences, goat erythrocyte being mammalian class, are biconcave in nature and possess similarities in basic properties to human erythrocytes. In addition, its easy availability has made goat erythrocyte a suitable study model in several studies [35–38]. Hence goat blood was used for our present study.