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Protective effect of ascorbic acid against oxidative damage induced by hydrogen peroxide in cultured human peripheral blood lymphocytes

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Abstract

Hydrogen peroxide is one of the reactive oxygen species for cellular injury. It is overproduced during oxidative stress and is known to damage proteins, nucleic acids and cell membranes. The present study was aimed to study the protective effect of ascorbic acid against the toxic doses of hydrogen peroxide using lipid peroxidation and cytokinesis blocked micronucleus assay. Hydrogen peroxide was studied at 50, 100 and 200μM and was found to increase a dose dependent increase in lipid peroxidation and micronuclei frequency. The treatment of 100 and 200μM of hydrogen peroxide separately along with 20, 40 and 80μM of ascorbic acid showed a dose dependent decrease in the lipid peroxidation and micronuclei frequency. The results suggest a protective effect of ascorbic acid against the hydrogen peroxide induced oxidative damage in cultured human peripheral blood lymphocytes.

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References

  1. Daroui P, Desai SO, Li TK, Liu AA, Liu LF. Hydrogen peroxide induces topoisomerase in mediated DNA Damage and cell death. J Biol Chem 2004; 279: 14587–14595.

    Article  PubMed  CAS  Google Scholar 

  2. Imlay JA, Chin SM, Linn S. DNA damage and oxygen radical toxicity. Science 1988; 240: 640–642.

    Article  PubMed  CAS  Google Scholar 

  3. Halliwell B, Gutteridge JM. Biologically relevant metal ion-dependent hydroxyl radical generation. An update. FEBS Lett 1992; 307: 108–112.

    Article  PubMed  CAS  Google Scholar 

  4. Josephson RA, Silverman HS, Lakatta EG, Stern MD, Zweier JL. Study of the mechanisms of Hydrogen peroxide and hydroxyl free radical induced cellular injury and calcium overload in cardiac myocytes. J Biol Chem 1991; 266: 2354–2361.

    PubMed  CAS  Google Scholar 

  5. Andreoli C, Leopardi P, Rossi S, Crebelli R. Processing of DNA damage induced by hydrogen peroxide and methyl methane sulfonate in human lymphocytes: analysis by alkaline single gel electrophoresis and cytogenetic methods. Mutagenesis 1999; 14: 497–503.

    Article  PubMed  CAS  Google Scholar 

  6. Pryor WA. Oxy radicals and related species, their formation, life times and reactions. Annu Rev Physiol 1986; 48: 657–667.

    Article  PubMed  CAS  Google Scholar 

  7. Birnboin HC. DNA strand breaks in human leukocytes induced by superoxide anion, hydrogen peroxide and tumor promoters are repaired slowly compared to breaks induced by ionizing radiation. Carcinogenesis 1986; 7: 1511–1517.

    Article  Google Scholar 

  8. Sofni T, Ishidate M. Induction of chromosomal aberrations in cultured Chinese hamsters cells superoxide generation system. Mutat Res 1984; 140: 27–31.

    Article  Google Scholar 

  9. Tsuda H. Chromosomal aberrations induced by hydrogen peroxide in cultured mammalian cells. Japanese J Genet 1981; 58: 1–8.

    Article  Google Scholar 

  10. Prise KM, Davies S, Michael BD. Cell killing and DNA damage in Chinese hamster V79 cells treated with hydrogen peroxide. Int J Radiat Biol 1989; 55: 583–592.

    Article  PubMed  CAS  Google Scholar 

  11. Thibodeau PA, Paquette B. DNA damage induced by catechol estrogens in the presence of copper (II): generation of reactive oxygen species and enhancement by NADH Free. Radic Biol Med 1999; 27: 1367–1377.

    Article  CAS  Google Scholar 

  12. Comporti M. Three models of free radical-induced cell injury. Chem Biol Interact 1989; 72: 1–56.

    Article  PubMed  CAS  Google Scholar 

  13. Eder E, Wacker M, Wanek P. Lipid peroxidation — related 1, N2-propandodeoxyguanosine-DNA adducts induced by endogenously formed 4-hydroxy-2-noneal in organs of female rats fed diets supplemented with sunflower, rapeseed live or coconut oil. Mutat Res 2004; 654: 101–107.

    Google Scholar 

  14. Esterbauer H, Schaur RJ, Zollner H. Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Rad Biol Med 1991; 11: 81–128.

    Article  PubMed  CAS  Google Scholar 

  15. Gerard-Monnier D, Erdelmeier I, Regnard K, Mozehenry N, Yadan JC, Chaudierel J. Reactions of 1-Methyl-2-phenylindole with malondialdehyde and 4-hydroxyalkenals Analytical applications to a colorimetric assay of lipid peroxidation. Chem Res Toxicol 1985; 11: 1176–1183.

    Article  Google Scholar 

  16. Carballo MA, Alvarez S, Boveris A. Cellular stress by light and Rose Bengal in human lymphocytes. Mutat Res 1993; 288: 215–222.

    PubMed  CAS  Google Scholar 

  17. Suryawanshi NP, Bhutey AK, Nagdeote AN, Jadhav AA, Manoorkar GS. Study of lipid peroxide and lipid profile in diabetes mellitus. Ind J Clin Biochem 2006; 27: 126–130.

    Article  Google Scholar 

  18. Fenech M, Morley AA. Measurement of micronuclei in lymphocytes. Mutat Res 1985; 147: 29–36.

    PubMed  CAS  Google Scholar 

  19. Fenech M, Chang WP, Krisch-Volders M, Holland SN, Bonassi S, Zeiger E. HUMN project: detailed description of the scoring criteria for the cytokinesis — block micronucleus assay using isolated human lymphocyte cultures. Mutat Res 2003; 534: 65–75.

    PubMed  CAS  Google Scholar 

  20. Ray G, Hussain SA. Oxidants, antioxidants and carcinogens. Ind J Exp Biol 2002; 40:1213–1232.

    CAS  Google Scholar 

  21. Hagmar L, Bonassi S, Stromberg U, Brogger A, Knudson JE, Norppa H, Reuterwall C. Chromosomal aberration in human lymphocytes predict human cancer: a report from the European study Group on Cytogenetic Biomarkers and Health (ESCH). Cancer Res 1998; 58: 4117–4121.

    PubMed  CAS  Google Scholar 

  22. Hagmar L, Brogger A, Hansteen IL, Heins S, Hoggstedt B, Knudsen L, et al. Cancer risk in humans predicted by increased level of chromosomal aberrations in human lymphocytes: Nordic study Group on the Health Risk of Chromosome Damge. Cancer Res 1994; 54: 2919–2922.

    PubMed  CAS  Google Scholar 

  23. Albertini RJ, Anderson D, Douglas GR, Hagmar L, Heminiki K, Merelo F, et al. IPCS guideline for the monitoring of genotoxic effects of carcinogens in humans. Mutat Res 2000; 463: 111–172.

    Article  PubMed  CAS  Google Scholar 

  24. Devipriya N, Sudheer AR, Srinivasan M, Menon VP. Quercetin ameliorates gamma radiation induced DNA damage and biochemical changes in human peripheral blood lymphocytes. Mutat Res 2008; 654: 1–7.

    PubMed  CAS  Google Scholar 

  25. Prasad NR, Srinivasan M, Pugalendi KV, Menon VP. Protective effect of ferfulic acid on γ-radiation induced micronuclei, dicentric aberration and lipid peroxidation in human lymphocytes. Mutat Res 2006; 603: 129–134.

    PubMed  CAS  Google Scholar 

  26. Singh M, Kaur P, Sandhir R, Kiran R. Protective effects of vitamin E against atrazine-induced genotoxicity in rats. Mutat Res 2008; 654: 145–149.

    PubMed  CAS  Google Scholar 

  27. Yousef MI, Awad TI, Elhag FA, Khaled FA. Study of the protective effect of ascorbic acid against the toxicity of stannous chloride on oxidative damage, anti-oxidant enzymes and biochemical parameters in rabbits. Toxicol 2007; 235: 194–202.

    Article  CAS  Google Scholar 

  28. Armutcu F, Coskun O, Gurel A, Sahin S, Kanter M, Cihan A, Numanoglu KV, Altinyazur C. Vitamin E protects against acetone induced oxidative stress in rat red blood cells. Cell Biol Toxicol 2005; 21: 53–60.

    Article  PubMed  CAS  Google Scholar 

  29. Bielski DHJ, Allen O, Schwarz HA. Mechanism of disproportionation of ascorbic radicals. J Am Chem Soc 1981; 103: 3516.

    Article  CAS  Google Scholar 

  30. Bielski BHJ. Chemistry of ascorbic acid radicals. In Seib, PA, Tolbert, B.M. Ed. Ascorbic acid: Chemistry, metabolism and uses. Washington, DC: American Chemical Society; 2002.

    Google Scholar 

  31. Szeto YT, Tomlinson B, Benzie IFF. Total antioxidant and ascorbic acid content of fresh fruits and vegetables: implications for dietary planning and food preservation. Br J Nutr 2002; 87: 55–59.

    Article  PubMed  CAS  Google Scholar 

  32. Edgar JA. Ascorbic acid and biological alkylating agents. Nature 1974; 248: 136–137.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Human Genetics and Toxicology Laboratory, Section of Genetics, Department of Zoology, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, 202002, U.P, India

    Yasir Hasan Siddique, Tanveer Beg & Mohammad Afzal

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  1. Yasir Hasan Siddique
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  2. Tanveer Beg
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  3. Mohammad Afzal
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Correspondence to Yasir Hasan Siddique.

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Siddique, Y.H., Beg, T. & Afzal, M. Protective effect of ascorbic acid against oxidative damage induced by hydrogen peroxide in cultured human peripheral blood lymphocytes. Indian J Clin Biochem 24, 294–300 (2009). https://doi.org/10.1007/s12291-009-0055-5

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