Mitochondrial damage as a mechanism of cell injury in the killing of cultured hepatocytes by tert-butyl hydroperoxide.
The killing of cultured hepatocytes by tert-butyl hydroperoxide (TBHP) occurs by different mechanisms depending on the presence or absence of the antioxidant N,N'-diphenylphenylenediamine (DPPD). In either situation there is evidence of mitochondrial damage. The mitochondrial inner membrane potential is lost, a result determined by the release from the cells of the lipophilic cation [3H]triphenylmethylphosphonium (TPMP+). Deenergization of the mitochondria is accompanied by a loss of ATP. Oligomycin reduced ATP stores without release of TPMP+ or without effect on the viability of the hepatocytes over the same time course that TBHP killed the majority of the cells. Monensin, a H+/Na+ ionophore, potentiated the toxicity of tert-butyl hydroperoxide in the presence or absence of DPPD. By contrast, extracellular acidosis reduced the toxicity of tert-butyl hydroperoxide in the presence or absence of DPPD. Neither monensin nor extracellular acidosis affected the metabolism of tert-butyl hydroperoxide, the release of TPMP+, or the extent of the peroxidation of cellular lipids. These data document the presence of mitochondrial damage in hepatocytes intoxicated with TBHP in both the presence and absence of DPPD. Furthermore, the potentiation by monensin is readily explained by the proposal that mitochondrial deenergization is accompanied by an intracellular acidosis. Such acidosis tends to delay the development of lethal cell injury. The protective effect of extracellular acidosis supports this interpretation.
Masaki
N
N
Department of Pathology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
Kyle
M E
ME
Serroni
A
A
Farber
J L
JL
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D008054:Lipid Peroxides / Q000096:biosynthesis; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000473:pathology; D008985:Monensin / Q000633:toxicity; D010545:Peroxides / Q000633:toxicity*; D010655:Phenylenediamines / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013997:Time Factors; D020122:tert-Butylhydroperoxide
D008054:Lipid Peroxides; D010545:Peroxides; D010655:Phenylenediamines; D000255:Adenosine Triphosphate; D008985:Monensin; D020122:tert-Butylhydroperoxide; C004845:N,N'-diphenyl-4-phenylenediamine