Rousarova 2015 Abstract MiPschool London 2015
|Estimation of ROS production in isolated mitochondria after treatment with an acetaminophen metabolite.|
Event: MiPschool London 2015
Acetaminophen (APAP) is a frequently used analgetic and antipyretic drug. After overdose, it may cause a number of pathophysiological processes that can even lead to acute liver and/or kidney failure. The cause of toxicity can be recognized in its metabolic activation but the entire mechanism of acetaminophen toxicity is still unknown. APAP is metabolized in hepatocytes through various pathways. The most important pathway acting in overdose is oxidation of APAP by cytochrome P450 to a substance, which is detoxified by reaction with glutathione [1,2].
We suppose that the metabolite of acetaminophen can also cause toxicity. Thus the main goal of our study was to assess a possible toxic effect of this metabolite in isolated mitochondria using detection of ROS production. We used CM-H2DCFDA molecular probe that is nonfluorescent until oxidized by ROS . We used isolated mitochondria from rat liver and from kidney cells treated with mitochondrial substrates and inhibitors to localize the site of ROS production. We proved that kidney mitochondria and mitochondria from rat liver treated with the acetaminophen metabolite produced ROS in significantly higher extent in comparison with controls. In 5 mM solution, ROS production in mitochondria isolated from rat liver was enhanced 9-fold and 3-fold in presence of glutamate and malate (i.e. complex I-related) and succinate and rotenone (i.e. complex II-related), respectively. Similar results were found in mitochondria isolated from kidney cells. The results support our hypothesis about the possible toxic effect of acetaminophen metabolite likely contributing to the overall toxicity.
Stress:Oxidative stress;RONS Organism: Rat Tissue;cell: Liver, Kidney Preparation: Isolated mitochondria
Pathway: N, S
Affiliations and Support
1-Dept Biol Biochem Sc, Univ Pardubice
2-Dept Analytical Chem, Univ Pardubice
3-Dept Physiology, Charles Univ in Prague, Czech Republic. - [email protected]
The study was supported by grant MZCR NT/14320.
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