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Genotoxicity of chloroquine in rat liver cells: Protective role of free radical scavengers

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Abstract

The genotoxic effect of chloroquine (CQ), a 4-aminoquinoline antimalarial drug was investigated in rat liver cells using the alkaline comet assay. Chloroquine (0–1000 μmol/L) significantly increased DNA strand breaks of rat liver cells dose-dependently. Rat liver cells exposed to CQ (100–500 μmol/L) and treated with endonuclease III and formamidopyrimidine-DNA glycosylase, the bacterial DNA repair enzymes that recognize oxidized pyrimidine and purine, respectively, showed greater DNA damage than those not treated with the enzymes, providing evidence that CQ induced oxidation of purines and pyrimidines. Treatment of cells with 5 mmol/L N-acetylcysteine, an intracellular reactive oxygen species (ROS) scavenger, and 100 μmol/L and 250 μmol/L deferoxamine, an established iron chelator, significantly decreased the CQ-induced strand breaks and base oxidation, respectively. Similarly, the formation of DNA strand breaks and oxidized bases was prevented by vitamin C (10 μmol/L) (a water-soluble antioxidant), quercetin (50 μmol/L) (an antioxidant flavonoid), and kolaviron (30 μmol/L and 90 μmol/L) (an antioxidant and a liver hepatoprotective phytochemical). The results indicate that the genotoxicity of CQ in rat liver cells might involve ROS and that free radical scavengers may elicit protective effects in these cells.

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Abbreviations

CQ:

chloroquine

DFO:

deferoxamine

DMSO:

dimethyl sulfoxide

Endo III:

endonuclease III

FCS:

fetal calf serum

FPG:

formamidopyrimidine glycosylase

MES:

4-morpholinoethane sulfonic acid

NAC:

N-acetylcysteine

ROS:

reactive oxygen species

SDS:

sodium dodecyl sulfate

References

  • Anderson D, Yu TW, Phillips BJ, Schmezer P. The effect of various antioxidants and other modifying agents on oxygen-radical-generated DNA damage in human lymphocytes in the COMET assay. Mutat Res. 1994;307:261–71.

    CAS  PubMed  Google Scholar 

  • Aruoma OI, Halliwell B, Hoey BM, Butler J. The antioxidant action of N-acetylcysteine: its reaction with hydrogen peroxide, hydroxyl radical, superoxide, and hypochlorous acid. Free Rad Biol Med. 1989;6:593–97.

    Article  CAS  PubMed  Google Scholar 

  • Blasiak J, Arabsk M, Pertynski T, Malecka-Panas E, Wozniak K, Drzewoski J. DNA damage in human colonic mucosa cells evoked by nickel and protective action of quercetin–involvements of free radicals? Cell Biol Toxicol. 2002;18:279–88.

    Article  CAS  PubMed  Google Scholar 

  • Blasiak J, Sikora A, Wozniak K, Drzewoski J. Genotoxicity of streptozotocin in normal and cancer cells and its modulation by free radical scavengers. Cell Biol Toxicol. 2004;20:83–96.

    Article  CAS  PubMed  Google Scholar 

  • Breman JG, Egan A, Keusch GT. The intolerable burden of malaria: a new look at the numbers. Am J Trop Med Hyg. 2001;64:11.

    Google Scholar 

  • Chatterjee T, Muhkopadhyay A, Khan KA, Giri AK. Comparative mutagenic and genotoxic effects of three antimalarial drugs, chloroquine, primaquine and amodiaquine. Mutagenesis. 1998;13:619–24.

    CAS  PubMed  Google Scholar 

  • Chen TH, Chang PC, Chang MC, Lin YF, Lee HM. Chloroquine induces the expression of inducible nitric oxide synthase in C6 glioma cells. Pharmacol Res. 2005;51:329–6.

    Article  CAS  PubMed  Google Scholar 

  • Collins AR. Oxidative DNA damage, antioxidants and cancer. Bioassays. 1999;21:238–46.

    Article  CAS  Google Scholar 

  • Collins AR, Duthie SJ, Dobson VL. Direct enzymatic detection of endogenous base damage in human lymphocyte DNA. Carcinogenesis. 1993;14:1733–35.

    CAS  PubMed  Google Scholar 

  • Collins AR, Dobson VL, Dusinska M, Kennedy G, Stetina R. The comet assay: what can it really tell us? Mutat Res. 1997;375:183–93.

    CAS  PubMed  Google Scholar 

  • De D, Krogstad FM, Cogswell FB, Krogstad DJ. Aminoquinolines that circumvent resistance in Plasmodium falciparum in vitro. Am J Trop Med Hyg. 1996;55:579–83.

    CAS  PubMed  Google Scholar 

  • Duthie SJ, Collins AR, Duthie GG, Dobson VL. Quercetin and myricetin protect against hydrogen peroxide-induced DNA damage (strand breaks and oxidized pyrimidines) in human lymphocytes. Mutat Res. 1997; 393:223–31.

    CAS  PubMed  Google Scholar 

  • Emerole GO, Thabrew MI. Changes in some rat hepatic microsomal components induced by prolonged administration of chloroquine. Biochem Pharmacol. 1983;32:3005–9.

    Article  CAS  PubMed  Google Scholar 

  • Evans MD, Podmore ID, Daly GJ, Pewrrett D, Lunec J, Herbert KE. Detection of purine lesions in cellular DNA using single cell electrophoresis with FPG protein. Biochem Soc Trans. 1995;23:434S.

    CAS  PubMed  Google Scholar 

  • Farombi EO, Emerole GO. Interference of common antimalarial drugs with some hepatic microsomal components and drug metabolism: potential implications for toxicity. S Afr J Sci. 1998;94:303–4.

    CAS  Google Scholar 

  • Farombi EO, Olowu BI, Emerole GO. Effect of three structurally related antimalarial drugs on liver microsomal components and lipid peroxidation in rats. Comp Biochem Physiol. 2000;126:217–24.

    CAS  Google Scholar 

  • Farombi EO, Adoro S, Uhunmwangho S. Antimalarial drugs exacerbate rat liver microsomal lipid peroxidation in the presence of oxidants. Biosci Rep. 2001;21:353–9.

    Article  CAS  PubMed  Google Scholar 

  • Farombi EO, Akanni OO, Emerole GO. Antioxidative and scavenging activities of kolaviron in vitro. Pharm Biol. 2002;40:107–16

    Article  CAS  Google Scholar 

  • Farombi EO, Shyntum YY, Emerole GO. Influence of chloroquine treatment and malaria infection on some enzymatic and non-enzymatic antioxidants in humans. Drug Chem Toxicol. 2003;26:59–71.

    Article  CAS  PubMed  Google Scholar 

  • Farombi EO, Moller P, Dragsted LO. Ex-vivo and in-vitro protective effects of kolaviron against oxygen-derived radical-induced DNA damage and oxidative stress in human lymphocytes and rat liver cells. Cell Biol Toxicol. 2004;20:71–82.

    Article  PubMed  Google Scholar 

  • Hartmann A, Speit G. Genotoxic effects of chemicals in the single cell gel (SCG) test with human blood cells in relation to the induction of sister-chromatid exchanges (SCE). Mutat Res. 1995;346: 49–56.

    Article  CAS  PubMed  Google Scholar 

  • Iwu MM. Antihepatotoxic constituents of Garcina kola seeds. Experientia. 1985;41:699–70.

    Article  CAS  PubMed  Google Scholar 

  • Magwere T, Naik YS, Hasler J. Effects of chloroquine on antioxidant enzymes in rat liver and kidney. Free Radic Biol Med. 1997;22: 321–27.

    Article  CAS  PubMed  Google Scholar 

  • Meister A, Anderson ME. Glutathone. Annu Rev Biochem. 1983;52: 711–60.

    Article  CAS  PubMed  Google Scholar 

  • Middleton E, Kandaswami C, Theoharides TC. The effects of plant flavonoids on mammalian cells: implications for inflammation, heart disease and cancer. Pharmacol Rev. 2000;52:673–751.

    CAS  PubMed  Google Scholar 

  • Mosuro AAO. Genotoxicity of antimalarial drugs 1: the 4-aminoquinolines. Biokemistri. 1997;7:9–18.

    CAS  Google Scholar 

  • Nakayama T, Yamada M, Osawa T, Kawakishi S. Suppression of active-oxygen-induced cytotoxicity by flavonoids. Biochem Pharmacol. 1993;45:265–7.

    Article  CAS  PubMed  Google Scholar 

  • Nappi AJ, Vass E. Comparative studies of enhanced iron-mediated production of hydroxyl radical by glutathione, cysteine, ascorbic acid and selected catechols. Biochim Biophys Acta. 1997;1336:295–1.

    CAS  PubMed  Google Scholar 

  • Obaseki-Ebor EE, Obasi EE. Aspects of chloroquine mutagenicity. Mutat Res. 1986;175:51–9.

    Article  Google Scholar 

  • Reitz M, Lorei W, Lettko M. DNA single strand breaks in human lymphocytes after therapy with antirheumatic drugs. Arzneimittelforschung/Drug Res. 1991;41(II):1141–4.

    CAS  Google Scholar 

  • Riccio ES, Lee PS, Winegar RA, Krogstad DJ, De D, Mirsalis JC. Genetic toxicology testing of the antimalarial drugs chloroquine and a new analog, AQ-13. Environ Mol Mutagen. 2001;38:69–79.

    Article  CAS  PubMed  Google Scholar 

  • Schupbach ME. Chemically induced transmissible abnormalities in sperm-head shape. Mutat Res. 1979; 68:41–9.

    Article  CAS  PubMed  Google Scholar 

  • Summer AP, Stauffer WM, Fischer PR. Pediatric malaria in the developing world. Semin Pediatr Infect Dis. 2005;16:105–15.

    Article  PubMed  Google Scholar 

  • Snyder RD, Arnone MR. Putative identification of functional interactions between DNA intercalating agents and topoisomerase II using the V79 in vitro micronucleus assay. Mutat Res. 2002;503:21–35.

    CAS  PubMed  Google Scholar 

  • Thein N, Moller P, Amtoft H, et al. A strong genotoxic effect in mouse skin of a single painting of coal tar in hairless mice and in MutaTM Mouse. Mutat Res. 2000;468:117–24.

    CAS  PubMed  Google Scholar 

  • Thomas SM, Silburn KA, MacPhee DG. Frameshift mutagenesis by chloroquine in Escherichia coli and Salmonella typhimurium. Mutat Res. 1987;192:233–7.

    Article  CAS  PubMed  Google Scholar 

  • Tice RR, Agurell E, Anderson D, et al. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen. 2000;25:206–21.

    Article  Google Scholar 

  • Toler SM. Oxidative stress plays an important role in the pathogenesis of drug-induced retinopathy. Exp Biol Med. 2004;229:607–15.

    CAS  Google Scholar 

  • Weitberg AB, Weitzman SA. The effect of vitamin C on oxygen radical-induced sister chromatid exchanges. Mutat Res. 1985;144:23–6.

    Article  CAS  PubMed  Google Scholar 

  • Zegura B, Lah TT, Filipic M. The role of reactive oxygen species in microcystin-LR-induced DNA. Toxicology. 2004;200:59– 68.

    Article  CAS  PubMed  Google Scholar 

Download references

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  1. Drug Metabolism and Toxicology Research Laboratories, Department of Biochemistry, College of Medicine, University of Ibadan, Nigeria

    E. O. Farombi

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  1. E. O. Farombi
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Farombi, E.O. Genotoxicity of chloroquine in rat liver cells: Protective role of free radical scavengers. Cell Biol Toxicol 22, 159–167 (2006). https://doi.org/10.1007/s10565-006-0173-2

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  • DOI: https://doi.org/10.1007/s10565-006-0173-2

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