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Mitochondrial dysfunction as a mediator of hippocampal apoptosis in a model of hepatic encephalopathy

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Abstract

In this study, we describe the presence of apoptosis, associated with a mitochondrial dysfunction in the hippocampus of animals in an experimental model defined as minimal hepatic encephalopathy (MHE). This experimental model was studied after 10 days of induced portal vein calibrated stricture, leading to portal hypertension and to a moderate hyperammonemia, without the presence of other evident central nervous system changes. The molecular mechanisms here proposed indicate the presence of apoptotic intrinsic pathways that point to hippocampal mitochondria as an important mediator of apoptosis in this experimental model. In this model of MHE, the presence of DNA fragmentation is documented by 2.3-times increased number of TUNEL-positive cells. These findings together with a higher ratio of the Bcl-2 family members Bax/Bcl-xL in the outer mitochondrial membrane of the MHE animals together with 11% of cytochrome c release indicate the presence of apoptosis in this experimental model. A detailed analysis of the hippocampal mitochondrial physiology was performed after mitochondrial isolation. The determination of the respiratory rate in the presence of malate plus glutamate and ADP showed a 45% decrease in respiratory control in MHE animals as compared with the sham group. A marked decrease of cytochrome oxidase (complex IV of the electron transport chain) was also observed, showing 46% less activity in hippocampal mitochondria from MHE animals. In addition, mitochondria from these animals showed less ability to maintain membrane potential (ΔΨ m) which was 13% lower than the sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the sham group. In addition, in vitro studies performed in mitochondria from sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apoptotic signaling in the presence of NH4 + and calcium.

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Abbreviations

ADP:

Adenosine diphosphate

Ca2+ :

Calcium

CsA:

Cyclosporine A

DiOC6:

3,3′dihexyloxocarbocyanine iodide

FCCP:

Carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone

FL-1:

Green fluorescence

HE:

Hepatic encephalopathy

H2O2 :

Hydrogen peroxide

MHE:

Minimal hepatic encephalopathy

MSH:

Mannitol–sacarose–Hepes

MPT:

Mitochondrial permeability transition

NH4 + :

Ammonia

PVS:

Portal vein stricture

RCR:

Respiratory control rate

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Acknowledgments

Authors acknowledge to the University of Buenos Aires, the National Scientific Council (Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET) and to the Promotion National Agency of Science and Technology (ANPCyT) for their financial support.

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Authors and Affiliations

  1. Laboratory of Free Radical Biology, School of Pharmacy and Biochemistry, University of Buenos Aires, Junin 956, C1113AAD, Buenos Aires, Argentina

    J. Bustamante, S. Lores-Arnaiz & A. Boveris

  2. Laboratory of Portal Hypertension, School of Pharmacy and Biochemistry, University of Buenos Aires, Junin 956, C1113AAD, Buenos Aires, Argentina

    S. Tallis, D. M. Roselló, A. Lemberg & J. C. Perazzo

  3. Pathology Centre, Pathology Department, School of Medicine, University of Buenos Aires, J Uriburu 950, C1114AAD, Buenos Aires, Argentina

    N. Lago

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  1. J. Bustamante
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  2. S. Lores-Arnaiz
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  3. S. Tallis
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  4. D. M. Roselló
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  5. N. Lago
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  6. A. Lemberg
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  7. A. Boveris
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  8. J. C. Perazzo
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Correspondence to J. Bustamante.

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Bustamante, J., Lores-Arnaiz, S., Tallis, S. et al. Mitochondrial dysfunction as a mediator of hippocampal apoptosis in a model of hepatic encephalopathy. Mol Cell Biochem 354, 231–240 (2011). https://doi.org/10.1007/s11010-011-0822-5

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  • DOI: https://doi.org/10.1007/s11010-011-0822-5

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