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Arsenic trioxide induced indirect and direct inhibition of glutathione reductase leads to apoptosis in rat hepatocytes

BioMetals volume 27, pages 483–494 (2014)Cite this article

Abstract

Glutathione reductase (GR) is an essential enzyme which maintains the reduced state of a cell. Therefore GR malfunction is closely associated with several disorders related to oxidative damage. The present study reports toxic manifestation of arsenic trioxide in respect of GR leading to apoptosis. Isolated rat hepatocytes exposed to arsenic trioxide were analyzed for GR expression and activity. Arsenic resulted in a time dependent inhibition of GR mediated by the superoxide anion. The cellular demand of functional enzyme is achieved by concomitant rise in gene expression. However, direct inhibition of GR by arsenic trioxide was also evident. Furthermore, arsenic induced free radical mediated inhibition of GR was found to be partially uncompetitive and associated with time dependent decrease in the substrate binding rate. Externalization of phosphatidylserine, nuclear degradation, apoptosis inducing factor leakage, apoptosome formation, caspase activation, DNA damage and break down of PARP suggest consequential induction of apoptosis due to inhibition of GR. The implication of GR was further established from the reduced rate of caspase activation in the arsenic trioxide treated cell, supplemented with complete and incomplete enzyme systems.

Graphical Abstract

Glutathione reductase activity is inhibited by binding of arsenic directly with the enzyme and also with the enzyme substrate complex. The mixed or partially uncompetitive mode of inhibition elicits arsenic binding with the GR–GSSG as well as GR–NADPH complex more effectively than with GR alone. Stimulated GR synthesis and gsr expression is concomitant with inhibited enzyme activity, however, reduction in GR efficiency eventually leads to caspase dependent apoptosis.

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Acknowledgments

AR is grateful to Council for Scientific and Industrial Research for a Senior Research Fellowship, SC gratefully acknowledges DST for a SRF (Project No SR/SO/AS-22/2008) and SM is grateful to University Grants Commission for financial support and SB acknowledges the National Academy of Sciences, India for the award of Senior Scientist, Platinum Jubilee Fellowship.

Conflict of interest

No competing financial interest exists.

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Author notes

  1. Atish Ray

    Present address: Immunobiology Group, Department of Zoology, University of Delhi, Delhi, India

Authors and Affiliations

  1. Environmental Toxicology Laboratory, Department of Zoology, Centre for Advanced Studies, Visva-Bharati (A Central University), Santiniketan, 731235, India

    Atish Ray, Sarmishtha Chatterjee, Sandip Mukherjee & Shelley Bhattacharya

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  1. Atish Ray
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  2. Sarmishtha Chatterjee
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Correspondence to Shelley Bhattacharya.

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Ray, A., Chatterjee, S., Mukherjee, S. et al. Arsenic trioxide induced indirect and direct inhibition of glutathione reductase leads to apoptosis in rat hepatocytes. Biometals 27, 483–494 (2014). https://doi.org/10.1007/s10534-014-9722-y

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  • DOI: https://doi.org/10.1007/s10534-014-9722-y

Keywords

  • Apoptosis
  • Glutathione reductase
  • NAC
  • Free radical
  • Nrf2