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Regulation of apoptosis by glutathione redox state in PC12 cells exposed simultaneously to iron and ascorbic acid

Mayo Hiroi1, Tohru Ogihara, Kazuya Hirano

  • 1Division of Neonatology, Department of Pediatrics, Osaka Medical College, 2-7, Daigaku-machi, Takatsuki, Osaka 569-8686, Japan. ped025@poh.osaka-med.ac.jp

Free Radical Biology & Medicine
|March 23, 2005
PubMed

Insights

Iron and ascorbic acid (AA) trigger cell death in infants with perinatal asphyxia. Antioxidants like alpha-tocopherol and glutathione (GSH) prevent this, highlighting a key mechanism in neuroprotection.

Area of Science:

  • Neuroscience
  • Biochemistry
  • Cell Biology

Background:

  • Elevated non-protein-bound iron (NPBI) and ascorbic acid (AA) levels are observed in the cerebrospinal fluid of infants experiencing perinatal asphyxia.
  • Perinatal asphyxia is a critical condition in newborns that can lead to significant neurological damage.

Purpose of the Study:

  • To elucidate the cytotoxic mechanisms of iron and ascorbic acid (AA) in neuronal cells.
  • To investigate the role of oxidative stress and glutathione (GSH) redox state in iron-AA induced apoptosis.

Main Methods:

  • PC12 cells were treated with ferrous sulfate (FeSO4) and AA.
  • Apoptosis was assessed, and protective effects of alpha-tocopherol and GSH ethyl ester were evaluated.
  • Levels of free radical damage markers, intracellular GSH/GSSG ratio, and half-cell reduction potential (Ehc) were measured.

Main Results:

  • FeSO4 and AA synergistically induced apoptosis in PC12 cells, preventable by antioxidants.
  • Exposure to AA and ferrous NPBI triggered apoptosis within hours, associated with increased markers of free radical damage.
  • Apoptosis correlated with a progressive decrease in intracellular GSH, an increase in GSSG, and a shift in Ehc to a critical oxidizing potential (-200 mV to -165 mV).

Conclusions:

  • Ferrous iron catalyzes hydroxyl radical generation and lipid peroxidation, potentiated by AA.
  • Depletion of intracellular GSH leads to a critical increase in Ehc, triggering or enhancing the apoptotic cascade.
  • These findings explain the Fe-AA cytotoxicity mechanism and suggest therapeutic targets for neuroprotection.

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