Cookies help us deliver our services. By using our services, you agree to our use of cookies. More information

Stankova 2010 Toxicol In Vitro

From Bioblast
Publications in the MiPMap
Staňková P, Kučera O, Lotková H, Roušar T, Endlicher R, Cervinková Z (2010) The toxic effect of thioacetamide on rat liver in vitro. Toxicol In Vitro 24:2097-2103.

» PMID: 20600801

Stankova Pavla, Kucera Otto, Lotkova Halka, Rousar Tomas, Endlicher Rene, Cervinkova Zuzana (2010) Toxicol In Vitro

Abstract: Thioacetamide (TAA) is a hepatotoxin frequently used for experimental purposes which produces centrilobular necrosis after a single dose administration. In spite of the fact that oxidative stress seems to play a very important role in the mechanism of TAA-induced injury, the effect of TAA on hepatocytes in primary culture with respect to the influence on mitochondria has yet to be verified. Hepatocytes were incubated for 24h in a medium containing TAA (0-70 mmol/l). Glutathione content (GSH/GSSG), reactive oxygen species and malondialdehyde formation were assessed as markers of cell redox state. Toxicity was determined by lactate dehydrogenase leakage and WST-1 assay. The functional capacity of hepatocytes was evaluated from albumin and urea production. Mitochondrial metabolism was assessed by measuring mitochondrial membrane potential and oxygen consumption. Our results show that a profound decrease in the GSH level in hepatocytes precedes a sharp rise in endogenous ROS production. ROS production correlates with an increase in lipoperoxidation. Mitochondria are affected by TAA secondarily as a consequence of oxidative stress. Oxidation of the NADH-dependent substrates of respiratory Complex I is significantly more sensitive to the toxic action of TAA than oxidation of the flavoprotein-dependent substrate of Complex II. Mitochondria can also maintain their membrane potential better when they utilize succinate as a respiratory substrate. It appears that GSH should be depleted below a certain critical level in order to cause a marked increase in lipid peroxidation. Mitochondrial injury can then occur and cell death develops. Keywords: Hepatotoxicity

O2k-Network Lab: CZ Hradec Kralove Cervinkova Z, CZ Pardubice Rousar T


Stress:Oxidative stress;RONS 

Tissue;cell: Liver  Preparation: Permeabilized cells  Enzyme: Complex I, Complex II;succinate dehydrogenase  Regulation: mt-Membrane potential, Fatty acid 

Pathway: N, S  HRR: Oxygraph-2k