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Neurochemical Research

, Volume 35, Issue 2, pp 336–342 | Cite as

Ethyl Pyruvate Inhibits Peroxynitrite-induced DNA Damage and Hydroxyl Radical Generation: Implications for Neuroprotection

  • Wei Chen
  • Zhenquan Jia
  • Hong Zhu
  • Kequan Zhou
  • Yunbo Li
  • Hara P. Misra
Original Paper

Abstract

Ethyl pyruvate (EP) has recently been reported to afford protection against neurodegenerative disorders. However, the mechanism underlying EP-mediated neuroprotection remains to be elucidated. Because peroxynitrite has been extensively implicated in the pathogenesis of various forms of neurodegenerative disorders via its cytotoxic effects, this study was undertaken to investigate whether the neuroprotective effect of EP is associated with inhibition of peroxynitrite-induced DNA strand breaks, a critical event leading to peroxynitrite elicited cytotoxicity. Incubation of φX-174 plasmid DNA with 3-morpholinosydnonimine (SIN-1), a peroxynitrite generator, led to the formation of both single- and double-stranded DNA breaks in a concentration- and time- dependent manner. The presence of EP (0.5–10 mM) was found to significantly inhibit SIN-1-induced DNA strand breaks in a concentration-dependent fashion. The consumption of oxygen induced by 250 μM SIN-1 was found to be decreased in the presence of EP (0.5–10 mM), indicating that EP might affect the auto-oxidation of SIN-1. It was observed that incubation of the plasmid DNA with authentic peroxynitrite caused significant DNA strand breaks, which could also be dramatically inhibited by EP (0.5–10 mM). EPR spectroscopy in combination with spin-trapping technique using 5,5-dimethylpyrroline-N- oxide (DMPO) as a spin trap demonstrated the formation of DMPO-hydroxyl radical adducts (DMPO-OH) from authentic peroxynitrite, and that EP at 0.5–10 mM inhibited the adduct signal in a concentration-dependent manner. Taken together, these results demonstrate for the first time that EP can inhibit peroxynitrite-mediated DNA damage and hydroxyl radical generation.

Keywords

Ethyl pyruvate Peroxynitrite DNA strand breaks Neuroprotection 

Abbreviations

DMPO

5,5-Dimethylpyrroline-N-oxide

DMPO-OH

DMPO-hydroxyl adduct

DMPO-OOH

DMPO-superoxide spin adduct

EPR

Electron paramagnetic resonance

EP

Ethyl pyruvate

SOD

Superoxide dismutase

ROS

Reactive oxygen species

RNS

Reactive nitrogen species

SIN-1

3-Morpholinosydnonimine

Notes

Acknowledgments

This work was supported by grants from the NIH grant R01HL071190, and Harvey Peters Research Center Foundation.

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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Division of Biomedical Sciences, Edward Via Virginia College of Osteopathic MedicineVirginia Tech Corporate Research CenterBlacksburgUSA
  2. 2.College of Food Science and Biological EngineeringZhejiang Gongshang UniversityHangzhouChina
  3. 3.Department of Food Science and TechnologyVirginia Polytechnic Institute and State UniversityBlacksburgUSA

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