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Oxidative damage of isolated rat hepatocytes by tert-butylhydroperoxide
Authors: Křiváková Pavla | Roušar Tomáš | Kučera Otto | Lotková Halka | Lábajová Anna | Červinková Zuzana | Drahota Zdeněk
Year: 2006
Type of publication: ostatní - přednáška nebo poster
Name of source: Proceedings from XX. Biochemický zjazd
Publisher name: Slovak Academy of Sciences, Institute of Molecular Physiology and Genetics
Place: Bratislava
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
cze Oxidative damage of isolated rat hepatocytes by tert-butylhydroperoxide Oxidative stress is one of the most important mechanisms through which hepatotoxins induce cell death. Tert-butylhydroperoxide (tBHP) has been widely used as a model compound to mimic the effect of oxidative stress in various cell types. The aim of our work was to characterize toxic injury of isolated rat hepatocytes induced by tBHP. Hepatocytes were isolated from male Wistar rats by two steps collagenase perfusion. A portion of cells was used for measurement of O2 consumption (Oxygraph Oroboros-2k) and for evaluation of mitochondrial membrane potential, MMP (TPP+). Remaining hepatocytes were cultivated in collagen coated Petri dishes. To estimate the rate of toxic injury we measured TBARS, LDH, MMP (Rho 123, JC-1), and GSH/GSSH (HPLC). Tert-butylhydroperoxide increases lipoperoxidation which precedes LDH leakage and decreases activity of respiratory Complex I and Complex II, MMP and GSH/GSSG. Respiratory Complex I activity is much more sensitive to the peroxidative action of tBHP than the activity of Complex II. We also found that the mechanism of the tBHP effect on mitochondrial membrane potential is dependent on respiratory substrates and we can supposed two different mechanisms. One of them is inhibition of respiratory complex I and the second one is mitochondrial permeability transition pore opening.
eng Oxidative damage of isolated rat hepatocytes by tert-butylhydroperoxide Oxidative stress is one of the most important mechanisms through which hepatotoxins induce cell death. Tert-butylhydroperoxide (tBHP) has been widely used as a model compound to mimic the effect of oxidative stress in various cell types. The aim of our work was to characterize toxic injury of isolated rat hepatocytes induced by tBHP. Hepatocytes were isolated from male Wistar rats by two steps collagenase perfusion. A portion of cells was used for measurement of O2 consumption (Oxygraph Oroboros-2k) and for evaluation of mitochondrial membrane potential, MMP (TPP+). Remaining hepatocytes were cultivated in collagen coated Petri dishes. To estimate the rate of toxic injury we measured TBARS, LDH, MMP (Rho 123, JC-1), and GSH/GSSH (HPLC). Tert-butylhydroperoxide increases lipoperoxidation which precedes LDH leakage and decreases activity of respiratory Complex I and Complex II, MMP and GSH/GSSG. Respiratory Complex I activity is much more sensitive to the peroxidative action of tBHP than the activity of Complex II. We also found that the mechanism of the tBHP effect on mitochondrial membrane potential is dependent on respiratory substrates and we can supposed two different mechanisms. One of them is inhibition of respiratory complex I and the second one is mitochondrial permeability transition pore opening. Oxidative stress, isolated hepatocytes, t-BHP, membrane potential

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