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Evaluation of Oxidative Stress in Acetaminophen Treated Hepatocytes In Vitro
Authors: Roušar Tomáš | Kučera Otto | Křiváková Pavla | Lotková Halka | Mužáková Vladimíra | Červinková Zuzana
Year: 2008
Type of publication: článek ve sborníku
Name of source: Toxicology Letters
Publisher name: Elsevier Ireland Ltd
Place: Shannon
Page from-to: 114
Titles:
Language Name Abstract Keywords
cze Zhodnocení oxidačního stresu v kultivovaných potkaních hepatocytů Zhodnocení oxidačního stresu v kultivovaných potkaních hepatocytů
eng Evaluation of Oxidative Stress in Acetaminophen Treated Hepatocytes In Vitro Acetaminophen (AAP) is one of the mostly used analgetics and antipyretics. It is considered to be a safe drug at therapeutic doses. In overdose, acetaminophen causes typical liver injury that may result in acute liver failure. Although a number of mechanisms contributing to the cell impairment have been described, the exact reason of the cell death remains unknown. Our purpose was to describe the AAP toxic acting in cultured rat hepatocytes using monitoring of ROS production, viability testing and glutathione assessment. The hepatocytes were isolated by two-steps collagenase reperfusion from male rats (Wistar, 240-280 g). Hepatocytes were incubated on collagenated Petri dishes or 24-well plates and treated with AAP dissolved in Williams´E medium (1, 2.5, 5, 10 and 20 mM AAP). Cells were incubated for following 24 hours and the biochemical changes were estimated. We assessed ROS production by specific ROS-probe (DCFDA), the viability was estimated by WST-1 test and LDH activity and as an antioxidant, the glutathione levels were measured. The increased levels of ROS production were found already after 3 h in three highest concentrations. At 12 h, the increase of ROS levels was: 80%; 200%; 410%; 490% and 780%, in 1, 2.5, 5, 10 and 20 mM AAP, respectively (compared to control). At 24 h, the enhanced ROS production remained unchanged. Cell viability was decreasing in dose dependent manner during whole time. We proved that AAP toxicity is associated with enhancement of ROS production. Acknowledgement: cultured hepatocytes, acetaminophen toxicity, glutathione.

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