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Dysregulation of autophagy in rat liver with mitochondrial DNA depletion induced by the nucleoside analogue zidovudine

  • Organ Toxicity and Mechanisms
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Abstract

The nucleoside reverse transcriptase inhibitor zidovudine (AZT), used in HIV infection treatment, induces mitochondrial DNA (mtDNA) depletion. A cause–effect relationship between mtDNA status alterations and autophagy has been reported. Both events are common in several liver diseases, including hepatocellular carcinoma. Here, we have studied autophagy activation in rat liver with mtDNA depletion induced by AZT administration in drinking water for 35 days. AZT at a concentration of 1 mg/ml, but not 0.5 mg/ml in the drinking water, decreased mtDNA levels in rat liver and extrahepatic tissues. In liver, mtDNA-encoded cytochrome c oxidase 1 protein levels were decreased. Although serum biomarkers of liver and kidney toxicity remained unaltered, β-hydroxybutyrate levels were increased in liver of AZT-treated rats. Moreover, autophagy was dysregulated at two levels: (i) decreased induction signalling of this process as indicated by increases in autophagy inhibitors activity (AKT/mTOR), and absence of changes (Beclin-1, Atg5, Atg7) or decreases (AMPK/ULK1) in the expression/activity of pro-autophagy proteins; and (ii) reduced autophagosome degradation as indicated by decreases in the lysosome abundance (LAMP2 marker) and the transcription factor TFEB controlling lysosome biogenesis. This resulted in increased autophagosome abundance (LC3-II marker) and accumulation of the protein selectively degraded by autophagy p62, and the transcription factor Nrf2 in liver of AZT-treated rats. Nrf2 was activated as indicated by the up-regulation of antioxidant target genes Nqo1 and Hmox-1. In conclusion, rat liver with AZT-induced mtDNA depletion presented dysregulations in autophagosome formation and degradation balance, which results in accumulation of these structures in parenchymal liver cells, favouring hepatocarcinogenesis.

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Acknowledgements

This study was supported by the Spanish “Instituto de Salud Carlos III” (Grant FIS PI15/00179 and PI16/00598) co-financed by European Regional Development Fund (ERDF), the Ministry of Science and Innovation, Spain (SAF2013-40620-R and SAF2016-75197-R), the “Junta de Castilla y León”, Spain (SA015U13 and SA063P17), the “Fundacion Mutua Madrileña”, Spain (Call 2015) and the “Fundación Memoria de D. Samuel Solórzano Barruso”, Spain (FS/8-2017). The group is member of the Network for Cooperative Research on Membrane Transport Proteins (REIT) and CIBERehd. The authors thank Emma Keck for editing the English.

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  1. Laboratory of Experimental Hepatology and Drug Targeting, Institute of Biomedical Research of Salamanca (IBSAL), CIBERehd, University of Salamanca, 37007, Salamanca, Spain

    Ana Santos-Llamas, Maria J. Monte, Jose J. G. Marin & Maria J. Perez

  2. Research Unit, University Hospital of Salamanca, Edificio Departamental (Lab. 129), Campus Miguel de Unamuno, 37007, Salamanca, Spain

    Maria J. Perez

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  1. Ana Santos-Llamas
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Correspondence to Maria J. Perez.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

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Santos-Llamas, A., Monte, M.J., Marin, J.J.G. et al. Dysregulation of autophagy in rat liver with mitochondrial DNA depletion induced by the nucleoside analogue zidovudine. Arch Toxicol 92, 2109–2118 (2018). https://doi.org/10.1007/s00204-018-2200-5

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  • DOI: https://doi.org/10.1007/s00204-018-2200-5

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