open access

Vol 48, No 1 (2010)
ORIGINAL PAPERS
Published online: 2010-06-10
Submitted: 2011-12-19
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alpha-Amanitin induced apoptosis in primary cultured dog hepatocytes.

Jan Magdalan, Alina Ostrowska, Aleksandra Piotrowska, Ilona Izykowska, Marcin Nowak, Agnieszka Gomułkiewicz, Marzena Podhorska-Okołów, Adam Szelag, Piotr Dziegiel
DOI: 10.2478/v10042-010-0010-6
·
Folia Histochem Cytobiol 2010;48(1):58-62.

open access

Vol 48, No 1 (2010)
ORIGINAL PAPERS
Published online: 2010-06-10
Submitted: 2011-12-19

Abstract

Amatoxin poisoning is caused by mushroom species belonging to the genera Amanita, Galerina and Lepiota with the majority of lethal mushroom exposures attributable to Amanita phalloides. High mortality rate in intoxications with these mushrooms is principally a result of the acute liver failure following significant hepatocyte damage due to hepatocellular uptake of amatoxins. A wide variety of amatoxins have been isolated; however, alpha-amanitin (alpha-AMA) appears to be the primary toxin. Studies in vitro and in vivo suggest that alpha-AMA does not only cause hepatocyte necrosis, but also may lead to apoptotic cell death. The objective of this study was to evaluate the complex hepatocyte apoptosis in alpha-AMA cytotoxicity. All experiments were performed on primary cultured canine hepatocytes. The cells were incubated for 12 h with alpha-AMA at a final concentration of 1, 5, 10 and 20 microM. Viability test (MTT assay), apoptosis evaluation (TUNEL reaction, detection of DNA laddering and electron microscopy) were performed at 6 and 12 h of exposure to alpha-AMA. There was a clear correlation between hepatocyte viability, concentration of alpha-AMA and time of exposure to this toxin. The decline in cultured dog hepatocyte viability during the exposure to alpha-AMA is most likely preceded by enhanced cellular apoptosis. Our results demonstrate that apoptosis might contribute to pathogenesis of the severe liver injury in the course of amanitin intoxication, particularly during the early phase of poisoning.

Abstract

Amatoxin poisoning is caused by mushroom species belonging to the genera Amanita, Galerina and Lepiota with the majority of lethal mushroom exposures attributable to Amanita phalloides. High mortality rate in intoxications with these mushrooms is principally a result of the acute liver failure following significant hepatocyte damage due to hepatocellular uptake of amatoxins. A wide variety of amatoxins have been isolated; however, alpha-amanitin (alpha-AMA) appears to be the primary toxin. Studies in vitro and in vivo suggest that alpha-AMA does not only cause hepatocyte necrosis, but also may lead to apoptotic cell death. The objective of this study was to evaluate the complex hepatocyte apoptosis in alpha-AMA cytotoxicity. All experiments were performed on primary cultured canine hepatocytes. The cells were incubated for 12 h with alpha-AMA at a final concentration of 1, 5, 10 and 20 microM. Viability test (MTT assay), apoptosis evaluation (TUNEL reaction, detection of DNA laddering and electron microscopy) were performed at 6 and 12 h of exposure to alpha-AMA. There was a clear correlation between hepatocyte viability, concentration of alpha-AMA and time of exposure to this toxin. The decline in cultured dog hepatocyte viability during the exposure to alpha-AMA is most likely preceded by enhanced cellular apoptosis. Our results demonstrate that apoptosis might contribute to pathogenesis of the severe liver injury in the course of amanitin intoxication, particularly during the early phase of poisoning.
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About this article
Title

alpha-Amanitin induced apoptosis in primary cultured dog hepatocytes.

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 48, No 1 (2010)

Pages

58-62

Published online

2010-06-10

DOI

10.2478/v10042-010-0010-6

Bibliographic record

Folia Histochem Cytobiol 2010;48(1):58-62.

Authors

Jan Magdalan
Alina Ostrowska
Aleksandra Piotrowska
Ilona Izykowska
Marcin Nowak
Agnieszka Gomułkiewicz
Marzena Podhorska-Okołów
Adam Szelag
Piotr Dziegiel

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