Publication detail

Analysis of oxidative/antioxidant balance in toxic liver injury.

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	Analysis of oxidative/antioxidant balance in toxic liver injury.	Introduction: Oxidative stress is one of the most important processes playing crucial role in toxic injury of the hepatocytes. Hepatotoxic injury may be caused by both increased levels of reactive oxygen species (ROS), and by decreased activity of antioxidant systems. Therefore, sufficient characterisation of oxidative stress requires use of various methods. These can help to distinguish particular components of the oxidative processes, but also antioxidant system. The aim of our work was to evaluate the oxidative stress and the changes in antioxidant capacity during toxic injury of isolated rat hepatocytes induced by model compound - tert-butylhydroperoxide (tBHP). Methods: Hepatocytes were isolated from male Wistar rats by two step collagenase perfusion. Isolated hepatocytes were cultivated in collagen coated Petri dishes. Mitochondria were isolated by differential centrifugation. The intracellular ROS probes, DCFDA (in both cell cultures, and isolated mitochondria) were used to measure the relation of different tBHP concentrations and ROS production. Levels of glutathione, as the main intracellular antioxidant, were analyzed together with the activity of glutathione reductase. As the marker of oxidative stress TBARS levels were measured. Preliminary conclusions: As supposed, we found that the action of tBHP increases oxidative stress through various mechanisms. ROS production has been increased only up to tBHP concentration of 1.5 mM in both cultivated cells, and isolated mitochondria. The GSH/GSSG ratio has been decreased and TBARS levels increased in dependence on the tBHP concentration.
eng	Analysis of oxidative/antioxidant balance in toxic liver injury.	Introduction: Oxidative stress is one of the most important processes playing crucial role in toxic injury of the hepatocytes. Hepatotoxic injury may be caused by both increased levels of reactive oxygen species (ROS), and by decreased activity of antioxidant systems. Therefore, sufficient characterisation of oxidative stress requires use of various methods. These can help to distinguish particular components of the oxidative processes, but also antioxidant system. The aim of our work was to evaluate the oxidative stress and the changes in antioxidant capacity during toxic injury of isolated rat hepatocytes induced by model compound - tert-butylhydroperoxide (tBHP). Methods: Hepatocytes were isolated from male Wistar rats by two step collagenase perfusion. Isolated hepatocytes were cultivated in collagen coated Petri dishes. Mitochondria were isolated by differential centrifugation. The intracellular ROS probes, DCFDA (in both cell cultures, and isolated mitochondria) were used to measure the relation of different tBHP concentrations and ROS production. Levels of glutathione, as the main intracellular antioxidant, were analyzed together with the activity of glutathione reductase. As the marker of oxidative stress TBARS levels were measured. Preliminary conclusions: As supposed, we found that the action of tBHP increases oxidative stress through various mechanisms. ROS production has been increased only up to tBHP concentration of 1.5 mM in both cultivated cells, and isolated mitochondria. The GSH/GSSG ratio has been decreased and TBARS levels increased in dependence on the tBHP concentration.	Oxidative stress, isolated hepatocytes, tBHP

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