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Early redox changes during rat thymocyte apoptosis

J Bustamante1, A Tovar-B, G Montero

  • 1National Laboratory of Free Radical Biology, School of Pharmacy and Biochemistry, University of Buenos Aires, Argentina.

Archives of Biochemistry and Biophysics
|January 1, 1997
PubMed
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Early redox changes occur during apoptosis induction in rat thymocytes. These changes, including glutathione alterations, are linked to the specific apoptosis-inducing agent, not always oxidative stress.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Toxicology

Background:

  • Apoptosis, or programmed cell death, is a critical biological process.
  • Redox homeostasis plays a significant role in cellular signaling and apoptosis.
  • Understanding early cellular events during apoptosis is crucial for disease research.

Purpose of the Study:

  • To investigate early redox changes during apoptosis induced by methylprednisolone, etoposide, and thapsigargin in rat thymocytes.
  • To determine if these early redox alterations are indicative of oxidative stress.

Main Methods:

  • Induction of apoptosis in rat thymocytes using methylprednisolone, etoposide, and thapsigargin.
  • Measurement of intracellular hydrogen peroxide (H2O2) levels using DCFH oxidation.
  • Quantification of thiobarbituric acid-reactive substances (TBARS) as a marker of lipid peroxidation.

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  • Assessment of intracellular glutathione (GSH) and oxidized glutathione (GSSG) levels.

    • Main Results:

    • All three agents induced apoptosis, confirmed by DNA laddering and fragmentation.
    • Methylprednisolone and thapsigargin increased intracellular H2O2, while etoposide did not.
    • Thapsigargin significantly increased TBARS, indicating lipid peroxidation.
    • Intracellular glutathione levels decreased in all treated groups, with varying effects on GSSG.

    Conclusions:

    • Early redox changes are a common feature of apoptosis induction in rat thymocytes.
    • The observed redox alterations are stimulus-dependent and do not uniformly indicate oxidative stress.
    • The specific pattern of redox changes provides insights into the mechanism of action of different apoptosis-inducing agents.