Effect of 5-nitroindole on adenylate energy charge, oxidative phosphorylation, and lipid peroxidation in rat hepatocytes.
5-Nitroindole (NI), a mutagenic nitroarene, was assayed for cytotoxic effects on rat hepatocytes. After incubation with 25-100 microM NI, the adenylate energy charge of the hepatocytes decreased significantly as a result of the decrease in ATP and the increase in AMP. ATP depletion correlated well with the effects of NI on mitochondrial electron transfer and energy transduction in hepatocytes. Thus, NI (a) inhibited the antimycin-sensitive hepatocyte respiration; (b) inhibited NADH oxidation by disrupted hepatocyte mitochondria; (c) inhibited L-malate-L-glutamate oxidation by ADP-supplemented mitochondria; (d) in the absence of ADP, stimulated the same substrates and also succinate oxidation by mitochondria; (e) released the latent ATPase activity of mitochondrial F1F0-ATP synthase; (f) shifted the redox level of reduced cytochromes (c + c1) and b towards the oxidized state; (g) inhibited NADH oxidation by disrupted mitochondria in the vicinity of the NADH-dehydrogenase flavoprotein; (h) inhibited Ca2+ uptake by mitochondria using L-malate-L-glutamate as an energy source; (i) inhibited valinomycin-induced, endogenously energized K+ uptake, with little effect on the ATP-induced uptake; and (j) inhibited the MgATP-dependent contraction of Ca(2+)-swollen mitochondria. NI inhibited lipid peroxidation in hepatocytes and also in substrate-supplemented liver microsomes and mitochondria, thus ruling out hydroperoxides as a cause of NI cytotoxicity. Long-term incubation with NI produced loss of hepatocyte viability, as indicated by lactate dehydrogenase leakage.
Dubin
M
M
Centro de Investigaciones Bioenergéticas, Facultad de Medicina, Paraguay, Buenos Aires, Argentina.
Carrizo
P H
PH
Biscardi
A M
AM
Fernandez Villamil
S H
SH
Stoppani
A O
AO
D000249:Adenosine Monophosphate / Q000737:chemistry*; D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D007211:Indoles / Q000633:toxicity*; D015227:Lipid Peroxidation / Q000187:drug effects*; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008930:Mitochondria, Liver / Q000737:chemistry / Q000187:drug effects; D009581:Nitrofurans / Q000494:pharmacology; D010085:Oxidative Phosphorylation / Q000187:drug effects*; D006180:Proton-Translocating ATPases / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013053:Spectrophotometry
D007211:Indoles; D009581:Nitrofurans; D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases; C090067:5-nitroindole; D002118:Calcium