Publikace detail

Model of acetaminophen-induced injury of rat hepatocytes in primary culture.

Autoři: Kučera Otto | Roušar Tomáš | Křiváková Pavla | Lotková Halka | Červinková Zuzana

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	Model of acetaminophen-induced injury of rat hepatocytes in primary culture.	Acetaminophen (AAP) overdose is the most common cause of druginduced acute liver failure. While metabolic activation of AAP is an essential step in the development of toxic lesion, precise mechanisms of the action of AAP in hepatotoxicity remain to be elucidated. The aim of our study was to establish a model of AAP-induced toxic injury of rat hepatocytes in vitro. Rat hepatocytes were incubated in supplemented William´s E medium without or with AAP (1 ? 10 mM) for 24 h. Toxicity of AAP was assessed at various time intervals up to 24 h by LDH activity in medium and by activity of mitochondrial dehydrogenases. All other measurements were performed after 24 h of incubation with AAP. Production of TBARS served as a marker of lipoperoxidation. Functional capacity of hepatocytes was evaluated by production of albumin. Contents of GSH and GSSG were assessed by HPLC. Morphological changes of cells were observed using phase contrast microscopy. Mitochondrial membrane potential (MMP) was evaluated using mitochondrial probe JC-1. Production of ROS was monitored by the fluorescence emission of DCFDA. AAP in dose- and time- dependent manner caused damage to hepatocyte membrane and decrease in mitochondrial dehydrogenases activity. Production of albumin, GSH content in hepatocytes and number of cells resp. mitochondria with high MMP were in reciprocal proportion to the dose of AAP after 24 h. Production of MDA increased with the dose of AAP. ROS production reached the highest level at AAP concentration of 3.75 mM. AAP in the range of 2.5 to 5 mM and 24h incubation are optimal for model injury to hepatocytes in vitro.
eng	Model of acetaminophen-induced injury of rat hepatocytes in primary culture.	Acetaminophen (AAP) overdose is the most common cause of druginduced acute liver failure. While metabolic activation of AAP is an essential step in the development of toxic lesion, precise mechanisms of the action of AAP in hepatotoxicity remain to be elucidated. The aim of our study was to establish a model of AAP-induced toxic injury of rat hepatocytes in vitro. Rat hepatocytes were incubated in supplemented William´s E medium without or with AAP (1 ? 10 mM) for 24 h. Toxicity of AAP was assessed at various time intervals up to 24 h by LDH activity in medium and by activity of mitochondrial dehydrogenases. All other measurements were performed after 24 h of incubation with AAP. Production of TBARS served as a marker of lipoperoxidation. Functional capacity of hepatocytes was evaluated by production of albumin. Contents of GSH and GSSG were assessed by HPLC. Morphological changes of cells were observed using phase contrast microscopy. Mitochondrial membrane potential (MMP) was evaluated using mitochondrial probe JC-1. Production of ROS was monitored by the fluorescence emission of DCFDA. AAP in dose- and time- dependent manner caused damage to hepatocyte membrane and decrease in mitochondrial dehydrogenases activity. Production of albumin, GSH content in hepatocytes and number of cells resp. mitochondria with high MMP were in reciprocal proportion to the dose of AAP after 24 h. Production of MDA increased with the dose of AAP. ROS production reached the highest level at AAP concentration of 3.75 mM. AAP in the range of 2.5 to 5 mM and 24h incubation are optimal for model injury to hepatocytes in vitro.	Oxidative stress, isolated hepatocytes, acetaminophen

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