Publikace detail

Oxidative damage of isolated rat hepatocytes by tert-butylhydroperoxide

Rok: 2006

Druh publikace: ostatní - přednáška nebo poster

Název zdroje: Proceedings from XX. Biochemický zjazd

Název nakladatele: Slovak Academy of Sciences, Institute of Molecular Physiology and Genetics

Místo vydání: Bratislava

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
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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