Effect of calcium and diltiazem on phenylhydrazine-induced oxidative injury in goat erythrocytes

Kaushik Das, Jharna Bhattacharyya

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DOI: 10.4236/health.2010.210181                

Abstract

Lipid peroxidation, hemolysis and thiol contents were studied in intact goat erythrocytes exposed to phenylhydrazine. An increase in lipid peroxidation, hemolysis and thiol content was observed after phenylhydrazine treatment of erythrocyte. Extracellular Ca2+ potentiates the phenylhydrazine-induced lipid peroxidation and hemoly- sis of erythrocytes significantly. Ca2+ does not influence much the thiol content of phenylhydrazine treated erythrocytes. No effect of Ca2+ on control lipid peroxidation, hemolysis and thiol contents of erythrocytes was observed. Diltiazem and EDTA inhibited the increased responses of lipid peroxidation and hemolysis due to Ca2+. However the thiol content was not much influenced by either diltiazem or EDTA. The results suggest that oxidative damage of erythrocyte caused by phenyl hydrazine could be prevented by calcium channel antagonist, diltiazem, which may act as antioxidant also.

Keywords

Ca2+; Diltiazem; Erythrocyte; Free Radical; Lipid Peroxidation; Phenylhydrazine

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Biswas, S., Bhattacharyya, J. and Datta, A.G. (2005) Ox- idant induced injury of erythrocyte-role of green tea leaf and ascorbic acid. Molecular and Cellular Biochemistry, 276(1-2), 205-210.
[2]	Latunde-Dada, G.O., Vulpe, C.D., Anderson, G.J., Sim- pson, R.J. and McKie, A.T. (2004) Tissue-specific chan- ges in iron metabolism genes in mice following phenylhydrazine-induced haemolysis. Biochimica et Biophysica Acta, 1690(2), 169-176.
[3]	Halder, J. and Bhaduri, A.N. (1998) Protective role of bla- ck tea against oxidative damage of human red blood cells. Biochem. Biophysical Research Communications, 244(3), 903-907.
[4]	Shetlar, M.D. and Hill, H.A. (1985) Reactions of hemog- lobin with phenylhydrazine: A review of selected aspects. Environmental Health Perspectives, 64, 265-281.
[5]	Bhattacharyya, J., Thompson, K.D. and Sayeed, M.M. (1993) Skeletal muscle Ca2+ flux and catabolic response during sepsis. American Journal of Physiology, 265(3), 487-493.
[6]	Braughler, J.M., Dyncan, L.A. and Goodman, T. (1985) Calcium enhances in vitro free radical induced damage to brain synaptosomes mitochondria and cultured spinal cord neurons. Journal of Neurochemistry, 45(4), 1288- 1293.
[7]	Lu, C., Chan, S.L., Fu, W. and Mattson, M.P. (2002) The lipid peroxidation product 4-Hydroxyneal facilitates op- ening of voltage dependent Ca2+ channels in neurons by increasing protein tyrosine phosphorylation. Journal of Biological Chemistry, 277(27), 24368-24375.
[8]	Buege, J.A. and Aust, S.D. (1978) Microsomal lipid per- oxidation. Methods in Enzymology, 52, 302-310.
[9]	Zhang, A., Zhu, Q.Y., Luk, Y.S., Ho, K.Y., Fung, K.P. and Chen, Z.Y. (1997) Inhibitory effect of jasmine green tea epicatechin isomers on free radical induced lysis of red blood cells. Life Sciences, 61(4), 383-394.
[10]	Owens, C.W. and Belcher, R.V. (1965) A colorimetric micro-method for the determination of glutathione. Biochemical Journal, 94(3), 705-711.
[11]	Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254.
[12]	Williamson, P., Puchulu, E., Westerman, M. and Sc- hlegel, R.A. (1990) Erythrocyte membrane abnormalities in sickle cell disease. Biotechnology and Applied Biochemistry, 12(5), 523-528.
[13]	Nash, G.B., Boghossian, S., Parmar, J., Dormandy, J.A. and Bevan, D. (1989) Alteration of the mechanical properties of sickle cells by repetitive deoxygenation: role of calcium and the effects of calcium blockers. Journal of British Society of Haematology, 72(2), 260-264.
[14]	Ohnishi, S.T., Horiuchi, K.Y., Horiuchi, K., Jurman, M.E. and Sadanaga, K.K. (1986) Nitrendipine, nifedipine, and verapamil inhibit the in vitro formation of irreversibly sickled cells. Pharmacology, 32(5), 248-256.
[15]	Rojstaczer, N. and Triggle, D.J. (1994) Calcium channel antagonists as antioxidants. Cardiovascular Drug Reviews, 12(1), 70-84.
[16]	Mason, R.P., Mak, I.T., Trumbore, M.W. and Mason, P.E. (1999) Antioxidant properties of Ca antagonists related to membrane biophysical interactions. American Journal of Cardiology, 84(4A), 16-22.
[17]	Chowdhury, T.D., Das, N., Chattopadhyay, A. and Datta, A.G. (1999) Effect of oxidative stress and erythropoietin on cytoskeletal protein and lipid organization in human erythrocytes. Polish Journal of Pharmacology & Pharmacy, 51(4), 341-350.
[18]	Gutteridge, J.M. and Halliwell, B. (1990) The measurement and mechanism of lipid peroxidation in biological systems. Trends in Biochemical Sciences, 15(4), 129- 135.
[19]	Watanabe, H., Kobayashi, A., Yamamoto, T., Suzuki, S., Hayashi, H. and Yamazaki, N. (1990) Alterations of human erythrocyte membrane fluidity by oxygen-derived free radicals and calcium. Free Radical Biology and Medicine, 8(6), 507-514.
[20]	Deuticke, B., Heller, K.B. and Haest, C.W.M. (1987) Progressive oxidative membrane damage in erythrocytes afterpulse treatment with t-butyl-hydroperoxide. Biochimica et Biophysica Acta, 899(1), 113-124.
[21]	Jain, S.K. (1985) In vivo externalization of phosphatidylserine and phosphatidylethanolamine in the membrane bilayer and hyper-coagulability by the lipid peroxidation of erythrocytes in rats. Journal of Clinical Investigation, 76(1), 281-286.
[22]	Zachary, S.B., Jason, D.M., David, J.J. and David, C.M. (2005) Primaquine-induced hemolytic anaemia: role of membrane lipid peroxidation and cytoskeletal protein alterations in the hemotoxicityof 5-hydroxyprimaquine. Journal of Pharmacology and Experimental Therapeutics, 314(2), 838-845.