Abstract
Oxidant and free radical-generating system were used to promote oxidative damage in erythrocytes. Among the oxidants used, phenylhydrazine represents one of the most investigated intracellular free radical-generating probes, which in the presence of haemoglobin autooxidises and give rise to hydroxyl radical, a marker for cellular damage. Erythrocyte, as a single cell, is a good model to be used for studying the haemolytic mechanism of anaemia. Our present investigations reveal increased lipid peroxidation of erythrocyte using phenylhydrazine as well as other oxygen-generating systems (hydrogen peroxide, iron with hydrogen peroxide). It has further been observed that not only lipid peroxidation, phenylhydrazine causes significant elevation in methemoglobin formation, catalase activity and turbidity, in the above system, which are the typical characteristics of haemolytic anaemia. However, exogenous administration of green tea leaf extract and ascorbic acid as natural antioxidants and free radical scavengers were shown to protect separately increased lipid peroxidation caused by phenylhydrazine, though the degree of protection is more in case of green tea leaf extract than ascorbic acid. Results suggest that oxidative damage in vivo due to haemolytic disease may be checked to some extent by using natural antioxidants. (Mol Cell Biochem 276: 205–210, 2005)
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References
De Groot H: Reactive oxygen species in tissue injury. Hepatogastroenterology 41: 328–332, 1994
Grace PA: Ischaemia-reperfusion injury. Br J Surg 81: 637–647, 1994
Chattopadhyay A, Daschowdhury T, Basu MK, Datta AG: Effect of Cu2+-ascorbic acid on lipid peroxidation, Mg2+-ATPase activity and spectrin of RBC membrane and reversal by erythropoietin. Mol Cell Biochem 118: 23–30, 1992
Chattopadhyay A, Biswas S, Bandopadhyay D, Sarkar C, Datta AG: Effect of isoproteronol on lipid peroxidation and antioxidant enzymes of myocardial tissue of mice and protection by quinidine. Mol Cell Biochem 245: 43–49, 2003
Bhattacharyya J, Biswas S, Datta AG: Mode of action of endotoxin: role of free radicals and antioxidants. Curr Med Chem 11: 359–368, 2004
Halliwell B: How to characterize an antioxidant: an update. Bochem Soc Symp 61: 73–101, 1995
Chattopadhyay A, Daschowdhury T, Bandopadhyay D, Datta AG: Protective effect of erythropoietin on the oxidative damage of erythrocyte membrane by hydroxyl radical. Biochem Pharmacol 59: 419–425, 2000
Daschowdhury T, Das N, Chattopadhyay A, Datta AG: Effect of oxidative stress and erythropoietin on cytoskeletal protein and lipid organization in human erythrocytes. Pol J Pharmacol 51: 341–350, 1999
Goldberg B, Stern A: The mechanism of oxidative hemolysis produced by phenylhydrazine. Mol Pharmacol 13: 832–839, 1977
Misra HP, Fridovich J: The oxidation of phenylhydrazine: superoxide and mechanism. Biochemistry 15: 681–687, 1976
Jain SK, Hochstein P: Generation of superoxide radicals by hydrazine-induced hemolytic anaemia. Biochem Biophys. Acta 586: 128–136, 1979
Nijveldt RJ, Nood EV, van Hoorn DFC, Boelens PG, Norren KV, van Leeuwen PAM: Flavonoids: a review of probable mechanism of action and potential applications. Am J Clin Nutr 74: 418–425, 2001
Youdim KA, Shukitt-Hale B, Mackinnon S, Kalt W, Joseph JA: Polyphenolics enhance red blood cell resistance to oxidative stress: in vitro and in vivo. Biochim Biophys Acta 1523: 117–122, 2000
Korkina LG, Afanas’ev IB: Antioxidant and chelating properties of flavonoids. Adv Pharmacol 38: 151–163, 1997
Hanasaki Y, Ogawa S, Fukui S: The correlation between active oxygens scavenging and antioxidative effects of flavonoids. Free Rad Biol Med 16: 845–850, 1994
Kerry NL, Abbey M: Red wine and fractionated phenolic compounds prepared from red wine inhibit low density lipoprotein oxidation in vitro. Atherosclerosis 135: 93–102, 1997
Karbownik M, Reiter RJ, Garcia JJ, Tan D-X: Melatonin reduces phenylhydrazine-induced oxidative damage to cellular membranes: evidence for the involvement of iron. Int J Biochem Cell Biol 32: 1045–1054, 2000
Buege JA, Aust SD: Microsomal lipid peroxidation. Methods Enzymol 52: 302–310, 1978.
Evelyn KA, Malloy HT: Microdetermination of oxyhemoglobin, methemoglobin and sulfhemoglobin in a single sample of blood. J Biol Chem 126: 655–662, 1938
Bates DA, Winterbourn CC: Haemoglobin denaturation, lipid peroxidation and hemolysis in phenylhydrazine-induced anaemia. Biochim Biophys Acta 798: 84–87, 1984
Beers RF(Jr), Sizer IW: A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195: 133–140, 1952
Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254, 1976
Clemens MR, Remmer H, Waller HD: Phenylhydrazine-induced lipid peroxidation of red blood cells in vitro and in vivo: monitoring by the production of volatile hydrocarbons. Biochem Pharmacol 33: 1715–1718, 1984
Goldstein BD, Rozen MG, Kunis RL: Role of red cell membrane lipid peroxidation in hemolysis due to phenylhydrazine. Biochem Pharmacol 29: 1355–1359, 1980
Schrier SL, Mohandas N: Globin chain specificity of oxidation-induced changes in red blood cell membrane properties. Blood 79: 1586–1592, 1992
Vinson J, Dabbagh Y, Serry M, Jang J: Plant flavoinoids, especially tea flavonols are powerful antioxidants using an in vitro oxidation model for heart disease. J Agri Food Chem 43: 2800–2802, 1995
Wei H, Zhang X, Zhao-Jt F, Wang Z-Y, Bickers D, Lebwohl M: Scavenging of hydrogen peroxide and inhibition of ultraviolet light induced oxidative DNA damage by aquous extracts from green and black teas. Free Rad Biol Med 26: 1427–1435, 1999
Sueoka N, Suganuma M, Sueoka E, Okabe S, Matsuyama S, Imai K, Nakachi K, Fujiki H: A new function of green tea: prevention of lifestyle-related diseases. Ann NY Acad Sci 928: 274–280, 2001
Sano J, Inami S, Seimiya K, Ohba T, Sakai S, Takano T, Mizuno K: Effects of green tea intake on the development of coronary artery disease. Circulation J 68: 665–670, 2004
Youdim KA, Shukilt-Hale B, Mackinnon S, Kalt W, Joseph JA: Polyphenolics enhance red blood cell resistance to oxidative stress in vitro and in vivo. Biochem Biophys Acta 1523: 117–122, 2000
Janero DR: Malondialdehyde and thiobarbituric acid reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Rad Biol Med 9: 515–540, 1990
Nandi A, Chatterjee IB: Scavenging of superoxide radical by ascorbic acid. J Bioscience 11: 435–441, 1987
Mukhopadhyay CK, Ghosh MK, Chatterjee IB: Ascorbic acid prevents lipid peroxidation and oxidative damage of proteins in guineapig extrahepatic tissue microsomes. Mol Cell Biochem 142: 71–78, 1995
Panda K, Chattopadhyay R, Ghosh MK, Chattopadhyay DJ, Chatterjee IB: Vitamin C prevents cigarette smoke induced oxidative damage of proteins and increased proteolysis. Free Rad Biol Med 27: 1064–1079, 1999
Frei B, England L, Ames BN: Ascorbate is an outstanding antioxidant in human blood plasma. Proc Natl Acad Sci 86: 6377–6381, 1989
Frei B: Ascorbic acid protects lipids in human plasma and low density lipoprotein against oxidative damage. Am J Clin Nutr 54(suppl.): 1113S–1118S, 1991
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Biswas, S., Bhattacharyya, J. & Dutta, A.G. Oxidant induced injury of erythrocyte—Role of green tea leaf and ascorbic acid. Mol Cell Biochem 276, 205–210 (2005). https://doi.org/10.1007/s11010-005-4062-4
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DOI: https://doi.org/10.1007/s11010-005-4062-4