Biochemical mechanism of irreversible cell injury caused by free radical-initiated reactions

Mol Cell Biochem. 1994 Aug 17;137(1):9-16. doi: 10.1007/BF00926034.


Effects of oxidative stress on isolated rat ventricular myocytes were studied. Myocyte viability was determined by the ability of these cells to retain rod-shaped morphology and to exclude trypan blue. The mean life time of myocytes was quantitated using the Weibull distribution function. Superfusion with 200 microM tert-butyl hydroperoxide (t-BHP) led to a time-dependent loss of cell viability, generation of the products of lipid peroxidation, oxidation of protein and non-protein thiols, a decrease in [ATP]i and in the cellular energy charge. Dithiothreitol (DTT, 5 mM) prolonged survival of myocytes exposed to t-BHP, attenuated oxidation of protein and non-protein thiols, and preserved the energy charge. Exposure to DTT did not affect the concentration of t-BHP-generated lipid peroxidation products. Promethazine (1 microM) prevented t-BHP-induced increase in the concentration of lipid peroxidation products, but did not prevent either loss of thiols or loss of cell viability. Superfusion with N-ethylmaleimide (NEM, 5 microM) also led to loss of cell viability, with accompanying decreases in protein and non-protein thiols, ATP and energy charge without the accumulation of the products of lipid peroxidation. Superfusion with FeSO4 (400 microM) and ascorbate (1 mM), (Fe-Asc) did not result in loss of cell viability or a decrease protein thiols or the energy charge. Superfusion with Fe-Asc, did, however, lead to a slight decrease in the concentration of non-protein thiols and ATP and a large increase in the concentration of lipid peroxidation products. Accumulation of lipid peroxidation products induced by Fe-Asc was prevented by promethazine.(ABSTRACT TRUNCATED AT 250 WORDS)

MeSH terms

  • Animals
  • Ascorbic Acid / pharmacology
  • Cell Survival
  • Ethylmaleimide / pharmacology
  • Ferrous Compounds / pharmacology
  • Free Radicals*
  • Lipid Peroxidation
  • Male
  • Myocardium / metabolism
  • Myocardium / pathology*
  • Oxidative Stress*
  • Peroxides / metabolism
  • Promethazine / pharmacology
  • Proteins / analysis
  • Rats
  • Rats, Sprague-Dawley
  • Thiobarbituric Acid Reactive Substances / analysis
  • tert-Butylhydroperoxide


  • Ferrous Compounds
  • Free Radicals
  • Peroxides
  • Proteins
  • Thiobarbituric Acid Reactive Substances
  • ferrous sulfate
  • tert-Butylhydroperoxide
  • Promethazine
  • Ethylmaleimide
  • Ascorbic Acid