Skip to main content
SpringerLink
Log in

Generation of superoxide from reaction of 3H-1,2-dithiole-3-thione with thiols: implications for dithiolethione chemoprotection

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

3H-1,2-Dithiole-3-thione (D3T), a potent member of dithiolethiones, induces phase 2 enzymes by activating an Nrf2/Keap1-dependent signaling pathway. It was proposed that interaction between D3T and two adjacent sulfhydryl groups of Keap1 might cause dissociation of Keap1 from Nrf2, leading to Nrf2 activation. This study was undertaken to investigate the reactions between D3T and thiols, including the dithiol compound, dithiothreitol (DTT), and the monothiol, glutathione (GSH). We reported here that under physiologically relevant conditions incubation of D3T with DTT caused remarkable oxygen consumption, indicating a redox reaction between D3T and the dithiol molecule. Incubation of D3T with GSH also led to oxygen consumption, but to a less extent. Electron paramagnetic resonance (EPR) studies showed that the redox reaction between D3T and DTT generated superoxide. Superoxide was also formed from the redox reaction of D3T with GSH. These findings demonstrate that D3T reacts with thiols, particularly a dithiol, generating superoxide, which may provide a mechanistic explanation for induction of Nrf2-dependent phase 2 enzymes by D3T.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

ARE:

Antioxidant response element

D3T:

3H-1,2-Dithiole-3-thione

DEPMPO:

5-(Diethoxyphosphoryl)-5-methylpyrroline N-oxide

DTPA:

Diethylenetriaminepentaacetic acid

DTT:

Dithiothreitol

EPR:

Electron paramagnetic resonance

GSH:

Glutathione

MAPKs:

Mitogen-activated protein kinases

NBT:

Nitroblue tetrazolium

PBS:

Phosphate-buffered saline

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

References

  1. Bailar JC 3rd, Gornik HL (1997) Cancer undefeated. N Engl J Med 336:1569–1574

    Article  PubMed  Google Scholar 

  2. Jacobson LP, Zhang BC, Zhu YR, Wang JB, Wu Y, Zhang QN, Yu LY, Qian GS, Kuang SY, Li YF, Fang X, Zarba A, Chen B, Enger C, Davidson NE, Gorman MB, Gordon GB, Prochaska HJ, Egner PA, Groopman JD, Munoz A, Helzlsouer KJ, Kensler TW (1997) Oltipraz chemoprevention trial in Qidong, People’s Republic of China: study design and clinical outcomes. Cancer Epidemiol Biomarkers Prev 6:257–265

    PubMed  CAS  Google Scholar 

  3. Kwak MK, Egner PA, Dolan PM, Ramos-Gomez M, Groopman JD, Itoh K, Yamamoto M, Kensler TW (2001) Role of phase 2 enzyme induction in chemoprotection by dithiolethiones. Mutat Res 480–481:305–315

    PubMed  Google Scholar 

  4. Nguyen T, Sherratt PJ, Pickett CB (2003) Regulatory mechanisms controlling gene expression mediated by the antioxidant response element. Annu Rev Pharmacol Toxicol 43:233–260

    Article  PubMed  CAS  Google Scholar 

  5. Motohashi H, Yamamoto M (2004) Nrf2-Keap1 defines a physiologically important stress response mechanism. Trends Mol Med 10:549–557

    Article  PubMed  CAS  Google Scholar 

  6. Li Y, Cao Z, Trush MA (2004) Chemical carcinogenesis and mutagenesis. In: F Barile (ed) Clinical toxicology: principles and mechanisms. CRC Press, Florida, pp 359–376

    Google Scholar 

  7. Kim W, Gates KS (1997) Evidence for thiol-dependent production of oxygen radicals by 4-methyl-5-pyrazinyl-3H-1,2-dithiole-3-thione (oltipraz) and 3H-1,2-dithiole-3-thione: possible relevance to the anticarcinogenic properties of 1,2-dithiole-3-thiones. Chem Res Toxicol 10:296–301

    Article  PubMed  CAS  Google Scholar 

  8. Velayutham M, Villamena FA, Fishbein JC, Zweier JL (2005) Cancer chemopreventive oltipraz generates superoxide anion radical. Arch Biochem Biophys 435:83–88

    Article  PubMed  CAS  Google Scholar 

  9. Velayutham M, Villamena FA, Navamal M, Fishbein JC, Zweier JL (2005) Glutathione-mediated formation of oxygen free radicals by the major metabolite of oltipraz. Chem Res Toxicol 18:970–975

    Article  PubMed  CAS  Google Scholar 

  10. Li Y, Kuppusamy P, Zweir JL, Trush MA (1996) Role of Cu/Zn-superoxide dismutase in xenobiotic activation. II. Biological effects resulting from the Cu/Zn-superoxide dismutase-accelerated oxidation of the benzene metabolite 1,4-hydroquinone. Mol Pharmacol 49:412–421

    PubMed  CAS  Google Scholar 

  11. Li Y, Zhu H, Kuppusamy P, Roubaud V, Zweier JL, Trush MA (1998) Validation of lucigenin (bis-N-methylacridinium) as a chemilumigenic probe for detecting superoxide anion radical production by enzymatic and cellular systems. J Biol Chem 273:2015–2023

    Article  PubMed  CAS  Google Scholar 

  12. Cao Z, Tsang M, Zhao H, Li Y (2003) Induction of endogenous antioxidants and phase 2 enzymes by alpha-lipoic acid in rat cardiac H9C2 cells: protection against oxidative injury. Biochem Biophys Res Commun 310:979–985

    Article  PubMed  CAS  Google Scholar 

  13. Frejaville C, Karoui H, Tuccio B, Le Moigne F, Culcasi M, Pietri S, Lauricella R, Tordo P (1995) 5-(Diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide: a new efficient phosphorylated nitrone for the in vitro and in vivo spin trapping of oxygen-centered radicals. J Med Chem 38:258–265

    Article  PubMed  CAS  Google Scholar 

  14. Shaked Z, Szajewski RP, Whitesides GM (1980) Rates of thiol-disulfide interchange reactions involving proteins and kinetic measurements of thiol pKa values. Biochemistry 19:4156–4166

    Article  PubMed  CAS  Google Scholar 

  15. Carey KA, Kensler TW, Fishbein JC (2001) Kinetic constraints for the thiolysis of 4-methyl-5-(pyrazin-2-yl)-1,2-dithiole-3-thione (oltipraz) and related dithiole-3-thiones in aqueous solution. Chem Res Toxicol 14:939–945

    Article  PubMed  CAS  Google Scholar 

  16. Delmas-Beauvieux MC, Peuchant E, Thomas MJ, Dubourg L, Pinto AP, Clerc M, Gin H (1998) The place of electron spin resonance methods in the detection of oxidative stress in type 2 diabetes with poor glycemic control. Clin Biochem 31:221–228

    Article  PubMed  CAS  Google Scholar 

  17. Timmins GS, Liu KJ, Bechara EJ, Kotake Y, Swartz HM (1999) Trapping of free radicals with direct in vivo EPR detection: a comparison of 5,5-dimethyl-1-pyrroline-N-oxide and 5-diethoxyphosphoryl-5-methyl-1-pyrroline-N-oxide as spin traps for HO· and SO4 ·-. Free Radic Biol Med 27:329–333

    Article  PubMed  CAS  Google Scholar 

  18. Stolze K, Udilova N, Nohl H (2000) Spin trapping of lipid radicals with DEPMPO-derived spin traps: detection of superoxide, alkyl and alkoxyl radicals in aqueous and lipid phase. Free Radic Biol Med 29:1005–1014

    Article  PubMed  CAS  Google Scholar 

  19. Ross D, Cotgreave I, Moldeus P (1985) The interaction of reduced glutathione with active oxygen species generated by xanthine-oxidase-catalyzed metabolism of xanthine. Biochim Biophys Acta 841:278–282

    PubMed  CAS  Google Scholar 

  20. Armstead WM (2001) Vasopressin induced cyclooxygenase dependent superoxide generation contributes to K+ channel function impairment after brain injury. Brain Res 910:19–28

    Article  PubMed  CAS  Google Scholar 

  21. Choi HS, Kim JW, Cha YN, Kim C (2006) A quantitative nitroblue tetrazolium assay for determining intracellular superoxide anion production in phagocytic cells. J Immunoassay Immunochem 27:31–44

    Article  PubMed  CAS  Google Scholar 

  22. Garrett IR, Boyce BF, Oreffo RO, Bonewald L, Poser J, Mundy GR (1990) Oxygen-derived free radicals stimulate osteoclastic bone resorption in rodent bone in vitro and in vivo. J Clin Invest 85:632–639

    Article  PubMed  CAS  Google Scholar 

  23. Nguyen T, Huang HC, Pickett CB (2000) Transcriptional regulation of the antioxidant response element: activation by Nrf2 and repression by MafK. J Biol Chem 275:15466–15473

    Article  PubMed  CAS  Google Scholar 

  24. Ishii T, Itoh K, Takahashi S, Sato H, Yanagawa T, Katoh Y, Bannai S, Yamamoto M (2000) Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages. J Biol Chem 275:16023–16029

    Article  PubMed  CAS  Google Scholar 

  25. Huang HC, Nguyen T, Pickett CB (2000) Regulation of the antioxidant response element by protein kinase C-mediated phosphorylation of NF-E2-related factor 2. Proc Natl Acad Sci USA 97:12475–12480

    Article  PubMed  CAS  Google Scholar 

  26. Itoh K, Wakabayashi N, Katoh Y, Ishii T, Igarashi K, Engel JD, Yamamoto M (1999) Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain. Genes Dev 13:76–86

    Article  PubMed  CAS  Google Scholar 

  27. Rosenberger J, Petrovics G, Buzas B (2001) Oxidative stress induces proorphanin FQ and proenkephalin gene expression in astrocytes through p38- and ERK-MAP kinases and NF-kappaB. J Neurochem 79:35–44

    Article  PubMed  CAS  Google Scholar 

  28. Schreck R, Albermann K, Baeuerle PA (1992) Nuclear factor kappa B: an oxidative stress-responsive transcription factor of eukaryotic cells. Free Radic Res Commun 17:221–237

    Article  PubMed  CAS  Google Scholar 

  29. Zhou LZ, Johnson AP, Rando TA (2001) NF kappa B and AP-1 mediate transcriptional responses to oxidative stress in skeletal muscle cells. Free Radic Biol Med 31:1405–1416

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported in part by NIH grant HL71190 (Y. L.). M. A. T. was supported by NIH grants ES03760, ES03819 and ES08078.

Author information

Authors and Affiliations

  1. Division of Biomedical Sciences, Edward Via Virginia College of Osteopathic Medicine, Virginia Tech Corporate Research Center, 2265 Kraft Drive, Blacksburg, VA, 24060, USA

    Zhenquan Jia, Hong Zhu, Hara P. Misra & Yunbo Li

  2. Division of Toxicology, Department of Environmental Health Sciences, The Johns Hopkins University Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD, 21205, USA

    Michael A. Trush

Authors
  1. Zhenquan Jia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hong Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael A. Trush
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hara P. Misra
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yunbo Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Hara P. Misra or Yunbo Li.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jia, Z., Zhu, H., Trush, M.A. et al. Generation of superoxide from reaction of 3H-1,2-dithiole-3-thione with thiols: implications for dithiolethione chemoprotection. Mol Cell Biochem 307, 185–191 (2008). https://doi.org/10.1007/s11010-007-9598-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-007-9598-z

Keywords

Search

Navigation