Stankova 2010 Toxicol In Vitro: Difference between revisions
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{{Publication | {{Publication | ||
|title=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. | |title=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. | ||
|info=[http://www.ncbi.nlm.nih.gov/pubmed/20600801 PMID: 20600801] | |info=[http://www.ncbi.nlm.nih.gov/pubmed/20600801 PMID: 20600801] | ||
|authors=Stankova | |authors=Stankova Pavla, Kucera Otto, Lotkova Halka, Rousar Tomas, Endlicher Rene, Cervinkova Zuzana | ||
|year=2010 | |year=2010 | ||
|journal=Toxicol In Vitro | |journal=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. | |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 | |keywords=Hepatotoxicity | ||
|mipnetlab=CZ Hradec Kralove Cervinkova Z, | |mipnetlab=CZ Hradec Kralove Cervinkova Z, CZ Pardubice Rousar T | ||
}} | }} | ||
{{Labeling | {{Labeling | ||
|tissues=Liver | |||
|preparations=Permeabilized cells | |||
|enzymes=Complex I, Complex II;succinate dehydrogenase | |||
|injuries=Oxidative stress;RONS | |||
|topics=mt-Membrane potential, Fatty acid | |||
|pathways=N, S | |||
|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
}} | }} | ||
Latest revision as of 10:58, 18 March 2020
| 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. |
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
Labels:
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
