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Hepatocytes isolated from preneoplastic rat livers are resistant to ethacrynic acid cytotoxicity

  • Organ Toxicity and Mechanisms
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Abstract

Glutathione S-transferases (GSTs) are involved in the detoxification of xenobiotics, such as several cytostatic drugs, through conjugation with glutathione (GSH). Pi class GST (GST P) liver expression is associated with preneoplastic and neoplastic development and contributes with the drug-resistance phenotype. Ethacrynic acid (EA) is an inhibitor of rat and human GSTs. In addition, causes lipid peroxidation in isolated rat hepatocytes. Therefore, we decided to evaluate the role of the GST/GSH system in isolated hepatocytes from preneoplastic rat livers (IP) in the presence of EA and determine the cytotoxicity of the drug. Our results showed a resistance to the toxic effects of EA since viability and cellular integrity values were significantly higher than control. Initial levels of thiobarbituric acid reactive substances (TBARS) in IP hepatocytes were significantly higher than control and the presence of EA did not change TBARS levels. A diminution in intracellular total GSH was observed by treating with EA isolated hepatocytes from both groups. However, the initial total GSH levels were higher in IP hepatocytes than in control. Immunoblotting analysis showed the presence of GST P in IP animals only. Although alpha and mu class isoenzymes levels were decreased in IP hepatocytes, total GST activity was 1.5-fold higher than in control. In addition, multidrug-resistance protein 2 (Mrp2) showed fivefold decreased levels in IP hepatocytes. In conclusion, increased total GSH, decreased Mrp2 levels and the presence of GST P could be critical factors involved in the resistance of IP hepatocytes to the toxicity of EA.

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Acknowledgments

Supported by research grants from Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) and from Consejo Nacional de Investigaciones Científicas y Técnicas. JPP is a recipient of a research fellowship from ANPCyT. We thank Dr. Aldo Mottino for the generous gift of the anti-Mrp2 antibody used in this study and Dr. Juan Monti and Elena Ochoa for their helpful technical assistance.

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  1. Facultad de Ciencias Bioquímicas y Farmacéuticas (UNR), Instituto de Fisiología Experimental (IFISE-CONICET), Suipacha 570, 2000, Rosario, Santa Fe, Argentina

    Juan Pablo Parody, María de Luján Alvarez, Ariel Quiroga, María Teresa Ronco, Daniel Francés, Cristina Carnovale & María Cristina Carrillo

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  1. Juan Pablo Parody
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  2. María de Luján Alvarez
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  3. Ariel Quiroga
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  4. María Teresa Ronco
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  5. Daniel Francés
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  6. Cristina Carnovale
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  7. María Cristina Carrillo
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Correspondence to María Cristina Carrillo.

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Parody, J.P., Alvarez, M.d.L., Quiroga, A. et al. Hepatocytes isolated from preneoplastic rat livers are resistant to ethacrynic acid cytotoxicity. Arch Toxicol 81, 565–573 (2007). https://doi.org/10.1007/s00204-007-0183-8

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  • DOI: https://doi.org/10.1007/s00204-007-0183-8

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