Publikace detail

Evaluation of Oxidative Stress in Acetaminophen Treated Hepatocytes In Vitro

Rok: 2008

Druh publikace: článek ve sborníku

Název zdroje: Toxicology Letters

Název nakladatele: Elsevier Ireland Ltd

Místo vydání: Shannon

Strana od-do: 114

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
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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