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Characterization of lapachol cytotoxicity: contribution of glutathione depletion for oxidative stress in Saccharomyces cerevisiae

  • Frederico A. V. Castro
  • Gabriel F. M. de Souza
  • Marcos D. PereiraEmail author
Original Article

Abstract

Over the years, quinones or its derivatives have been extensively studied due to their broad therapeutic spectrum. However, due to the significant structural differences between the individual naturally occurring quinones, investigation of the precise mechanism of their action is essential. In this context, we have analyzed the mechanism of lapachol [4-hydroxy-3-(3-methylbut-2-enyl)naphthalene-1,2-dione] toxicity using Saccharomyces cerevisiae as eukaryotic model organism. Analyzing yeast (wild type, sod1∆, and gsh1∆) cell growth, we observed a strong cytostatic effect caused by lapachol exposure. Moreover, survival of cells was affected by time- and dose-dependent manner. Interestingly, sod1∆ cells were more prone to lapachol toxicity. In this sense, mitochondrial functioning of sod1∆ cells were highly affected by exposure to this quinone. Lapachol also decreased glutathione (GSH) levels in wild type and sod1∆ cells even though glutathione disulfide (GSSG) remained unchanged. We believe that reduction of GSH contents has contributed to the enhancement of lipid peroxidation and intracellular oxidation, effect much more pronounced in sod1∆ cells. Overall, the collected data suggest that although lapachol can act as an oxidant, it seems that the main mechanism of its action initially consists in alkylation of intracellular targets such as GSH and then generating oxidative stress.

Notes

Funding information

This study received financial support from the Brazilian funding institutions CAPES, CNPq, and FAPERJ that allowed this work to be carried out.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

References

  1. Barclay BJ, DeHaan CL, Hennig UGG et al (2001) A rapid assay for mitochondrial DNA damage and respiratory chain inhibition in the yeast Saccharomyces cerevisiae. Environ Mol Mutagen 38:153–158.  https://doi.org/10.1002/em.1066 CrossRefGoogle Scholar
  2. Bernardi G (1979) The petite mutation in yeast. Trends Biochem Sci 4:197–201CrossRefGoogle Scholar
  3. Bernt E, Bergmeyer HU (1974) Glutathione. In: Methods of enzymatic analysis (Second English Edition). pp 1643–1647Google Scholar
  4. Bolton JL, Trush MA, Penning TM, Dryhurst G, Monks TJ (2000) Role of quinones in toxicology. Chem Res Toxicol 13:135–160CrossRefGoogle Scholar
  5. Castro FAV, Herdeiro RS, Panek AD, Eleutherio ECA, Pereira MD (2007) Menadione stress in Saccharomyces cerevisiae strains deficient in the glutathione transferases. Biochim Biophys Acta Gen Subj 1770:213–220.  https://doi.org/10.1016/j.bbagen.2006.10.013 CrossRefGoogle Scholar
  6. Castro FAV, Mariani D, Panek AD, Eleutherio ECA, Pereira MD (2008) Cytotoxicity mechanism of two naphthoquinones (menadione and plumbagin) in Saccharomyces cerevisiae. PLoS One 3:e3999.  https://doi.org/10.1371/journal.pone.0003999 CrossRefGoogle Scholar
  7. Costa WF, de Oliveira AB, Nepomuceno JC (2010) Genotoxicity of lapachol evaluated by wing spot test of Drosophila melanogaster. Genet Mol Biol 33:558–563.  https://doi.org/10.1590/S1415-47572010005000070 CrossRefGoogle Scholar
  8. Day M (2013) Yeast petites and small colony variants. In: Advances in applied microbiology, pp 1–41Google Scholar
  9. de Sá RA, de Castro FAV, Eleutherio ECA et al (2013) Brazilian propolis protects Saccharomyces cerevisiae cells against oxidative stress. Braz J Microbiol 44:993–1000.  https://doi.org/10.1590/S1517-83822013000300050 CrossRefGoogle Scholar
  10. Eckert KG, Eyer P, Sonnenbichler J, Zetl I (1990) Activation and detoxication of aminophenols. II. Synthesis and structural elucidation of various thiol addition products of 1,4-benzoquinoneimine and n-acetyl-1,4-benzoquinoneimine. Xenobiotica 20:333–350.  https://doi.org/10.3109/00498259009046851 CrossRefGoogle Scholar
  11. Elashvilis I, Elashvilis I, Butler E et al (1992) Yeast lacking superoxide dismutase. Mol Biol 2:18298–18302Google Scholar
  12. Esteves-Souza A, Figueiredo DV, Esteves A, Câmara CA, Vargas MD, Pinto AC, Echevarria A (2007) Cytotoxic and DNA-topoisomerase effects of lapachol amine derivatives and interactions with DNA. Braz J Med Biol Res 40:1399–1402.  https://doi.org/10.1590/S0100-879X2006005000159 CrossRefGoogle Scholar
  13. Esteves-Souza A, Araújo Lúcio K, da Cunha AS et al (2008) Antitumoral activity of new polyamine-naphthoquinone conjugates. Oncol Rep 20:225–231.  https://doi.org/10.3892/or.20.1.225 Google Scholar
  14. Evans MD, Dizdaroglu M, Cooke MS (2004) Oxidative DNA damage and disease: induction, repair and significance. Mutat Res Rev Mutat Res 567:1–61CrossRefGoogle Scholar
  15. Ferguson LR, von Borstel RC (1992) Induction of the cytoplasmic “petite” mutation by chemical and physical agents in Saccharomyces cerevisiae. Mutat Res Fundam Mol Mech Mutagen 265:103–148.  https://doi.org/10.1016/0027-5107(92)90042-Z CrossRefGoogle Scholar
  16. Fernandes PN, Mannarino SC, Silva CG, Pereira MD, Panek AD, Eleutherio ECA (2007) Oxidative stress response in eukaryotes: effect of glutathione, superoxide dismutase and catalase on adaptation to peroxide and menadione stresses in Saccharomyces cerevisiae. Redox Rep 12:236–244.  https://doi.org/10.1179/135100007X200344 CrossRefGoogle Scholar
  17. Fridovich I (1972) Superoxide radical and superoxide dismutase. Acc Chem Res 5:321–326.  https://doi.org/10.1021/ar50058a001 CrossRefGoogle Scholar
  18. Friedman JR, Nunnari J (2014) Mitochondrial form and function. Nature 505:335–343CrossRefGoogle Scholar
  19. Giulivi C, Boveris A, Cadenas E (1995) Hydroxyl radical generation during mitochondrial electron-transfer and the formation of 8-hydroxydesoxyguanosine in mitochondrial-DNA. Arch Biochem Biophys 316:909–916.  https://doi.org/10.1006/abbi.1995.1122 CrossRefGoogle Scholar
  20. Gómez Castellanos JR, Prieto JM, Heinrich M (2009) Red Lapacho (Tabebuia impetiginosa)-a global ethnopharmacological commodity? J Ethnopharmacol 121:1–13CrossRefGoogle Scholar
  21. Goulart MOF, Falkowski P, Ossowski T, Liwo A (2003) Electrochemical study of oxygen interaction with lapachol and its radical anions. Bioelectrochemistry 59:85–87.  https://doi.org/10.1016/S1567-5394(03)00005-7 CrossRefGoogle Scholar
  22. Grant CM, MacIver FH, Dawes IW (1997) Mitochondrial function is required for resistance to oxidative stress in the yeast Saccharomyces cerevisiae. FEBS Lett 410:219–222.  https://doi.org/10.1016/S0014-5793(97)00592-9 CrossRefGoogle Scholar
  23. Gutteridge JMC (1985) Superoxide dismutase inhibits the superoxide-driven Fenton reaction at two different levels. Implications for a wider protective role. FEBS Lett 185:19–23.  https://doi.org/10.1016/0014-5793(85)80732-8 CrossRefGoogle Scholar
  24. Hussain H, Green IR (2017) Lapachol and lapachone analogs: a journey of two decades of patent research (1997-2016). Expert Opin Ther Pat 27:1111–1121CrossRefGoogle Scholar
  25. Hussain H, Krohn K, Ahmad VU et al (2007) Lapachol: an overview. Arkivoc 2007:145–171.  https://doi.org/10.3998/ark.5550190.0008.204 CrossRefGoogle Scholar
  26. Inbaraj JJ, Chignell CF (2004) Cytotoxic action of juglone and plumbagin: a mechanistic study using HaCaT keratinocytes. Chem Res Toxicol 17:55–62.  https://doi.org/10.1021/tx034132s CrossRefGoogle Scholar
  27. Inoue M, Sato EF, Nishikawa M, Park AM, Kira Y, Imada I, Utsumi K (2005) Mitochondrial generation of reactive oxygen species and its role in aerobic life. Curr Med Chem 10:2495–2505.  https://doi.org/10.2174/0929867033456477 CrossRefGoogle Scholar
  28. Kalyanaraman B, Premovic PI, Sealy RC (1987) Semiquinone anion radicals from addition of amino acids, peptides, and proteins to quinones derived from oxidation of catechols and catecholamines. An ESR spin stabilization study. J Biol Chem 262:11080–11087Google Scholar
  29. Kucsera J, Yarita K, Takeo K (2000) Simple detection method for distinguishing dead and living yeast colonies. J Microbiol Methods 41:19–21.  https://doi.org/10.1016/S0167-7012(00)00136-6 CrossRefGoogle Scholar
  30. Lau SS, Hill BA, Highet RJ, Monks TJ (1988) Sequential oxidation and glutathione addition to 1,4-benzoquinone: correlation of toxicity with increased glutathione substitution. Mol Pharmacol 34:829–836Google Scholar
  31. Longo VD, Gralla EB, Valentine JS (1996) Superoxide dismutase activity is essential for stationary phase survival in Saccharomyces cerevisiae: mitochondrial production of toxic oxygen species in vivo. J Biol Chem 271:12275–12280.  https://doi.org/10.1074/jbc.271.21.12275 CrossRefGoogle Scholar
  32. Lu J-J, Bao J-L, Wu G-S, Xu WS, Huang MQ, Chen XP, Wang YT (2013) Quinones derived from plant secondary metabolites as anti-cancer agents. Anti Cancer Agents Med Chem 13:456–463.  https://doi.org/10.2174/1871520611313030008 Google Scholar
  33. Maeda M, Murakami M, Takegami T, Ota T (2008) Promotion or suppression of experimental metastasis of B16 melanoma cells after oral administration of lapachol. Toxicol Appl Pharmacol 229:232–238.  https://doi.org/10.1016/j.taap.2008.01.008 CrossRefGoogle Scholar
  34. Maistro EL, Fernandes DM, Pereira FMV, Andrade SF (2010) Lapachol induces clastogenic effects in rats. Planta Med 76:858–862.  https://doi.org/10.1055/s-0029-1240816 CrossRefGoogle Scholar
  35. Mauzeroll J, Bard AJ, Owhadian O, Monks TJ (2004) Menadione metabolism to thiodione in hepatoblastoma by scanning electrochemical microscopy. Proc Natl Acad Sci 101:17582–17587.  https://doi.org/10.1073/pnas.0407613101 CrossRefGoogle Scholar
  36. Monks TJ, Jones DC (2002) The metabolism and toxicity of quinones, quinonimines, quinone methides, and quinone-thioethers. Curr Drug Metab 3:425–438.  https://doi.org/10.2174/1389200023337388 CrossRefGoogle Scholar
  37. Oliveira MF, Lemos TLG, De Mattos MC et al (2002) New enamine derivatives of lapachol and biological activity. An Acad Bras Cienc 74:211–221.  https://doi.org/10.1590/S0001-37652002000200004 CrossRefGoogle Scholar
  38. Oliveira RAS, Azevedo-Ximenes E, Luzzati R, Garcia RC (2010) The hydroxy-naphthoquinone lapachol arrests mycobacterial growth and immunomodulates host macrophages. Int Immunopharmacol 10:1463–1473.  https://doi.org/10.1016/j.intimp.2010.08.023 CrossRefGoogle Scholar
  39. Pereira MD, Eleutherio ECA, Panek AD (2001) Acquisition of tolerance against oxidative damage in Saccharomyces cerevisiae. BMC Microbiol 1:1–10.  https://doi.org/10.1186/1471-2180-1-11 CrossRefGoogle Scholar
  40. Pereira MD, Herdeiro RS, Fernandes PN, Eleutherio ECA, Panek AD (2003) Targets of oxidative stress in yeast sod mutants. Biochim Biophys Acta Gen Subj 1620:245–251.  https://doi.org/10.1016/S0304-4165(03)00003-5 CrossRefGoogle Scholar
  41. Reddi AR, Culotta VC (2013) SOD1 integrates signals from oxygen and glucose to repress respiration. Cell 152:224–235.  https://doi.org/10.1016/j.cell.2012.11.046 CrossRefGoogle Scholar
  42. Ribeiro TP, Fernandes C, Melo KV, Ferreira SS, Lessa JA, Franco RWA, Schenk G, Pereira MD, Horn A Jr (2015) Iron, copper, and manganese complexes with in vitro superoxide dismutase and/or catalase activities that keep Saccharomyces cerevisiae cells alive under severe oxidative stress. Free Radic Biol Med 80:67–76.  https://doi.org/10.1016/j.freeradbiomed.2014.12.005 CrossRefGoogle Scholar
  43. Rodriguez CE, Shinyashiki M, Froines J, Yu RC, Fukuto JM, Cho AK (2004) An examination of quinone toxicity using the yeast Saccharomyces cerevisiae model system. Toxicology 201:185–196.  https://doi.org/10.1016/j.tox.2004.04.016 CrossRefGoogle Scholar
  44. Ross D, Thor H, Orrenius S, Moldeus P (1985) Interaction of menadione (2-methyl-1,4-naphthoquinone) with glutathione. Chem Biol Interact 55:177–184.  https://doi.org/10.1016/S0009-2797(85)80126-5 CrossRefGoogle Scholar
  45. Seung SA, Lee JY, Lee MY, Park JS, Chung JH (1998) The relative importance of oxidative stress versus arylation in the mechanism of quinone-induced cytotoxicity to platelets. Chem Biol Interact 113:133–144.  https://doi.org/10.1016/S0009-2797(98)00024-6 CrossRefGoogle Scholar
  46. Steels EL, Learmonth RP, Watson K (1994) Stress tolerance and membrane lipid unsaturation in Saccharomyces cerevisiae grown aerobically or anaerobically. Microbiology 140(Pt 3):569–576.  https://doi.org/10.1099/00221287-140-3-569 CrossRefGoogle Scholar
  47. Tsang CK w, Liu Y, Thomas J et al (2014) Superoxide dismutase 1 acts as a nuclear transcription factor to regulate oxidative stress resistance. Nat Commun 5:3446.  https://doi.org/10.1038/ncomms4446 CrossRefGoogle Scholar
  48. Wang CCC, Chiang Y-M, Sung S-C, Hsu YL, Chang JK, Kuo PL (2008) Plumbagin induces cell cycle arrest and apoptosis through reactive oxygen species/c-Jun N-terminal kinase pathways in human melanoma A375.S2 cells. Cancer Lett 259:82–98.  https://doi.org/10.1016/j.canlet.2007.10.005 CrossRefGoogle Scholar
  49. Ządzinski R, Fortuniak A, Bartosz G, Bilinski T, Grey M (2007) Menadione toxicity in Saccharomyces cerevisiae cells: activation by conjugation with glutathione. IUBMB Life 44:747–759.  https://doi.org/10.1080/15216549800201792 CrossRefGoogle Scholar

Copyright information

© Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2019

Authors and Affiliations

  1. 1.Departamento de Bioquímica, Instituto de QuímicaUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil

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