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Mitochondrial dysfunction as a mediator of hippocampal apoptosis in a model of hepatic encephalopathy
J Bustamante1, S Lores-Arnaiz, S Tallis
1Laboratory of Free Radical Biology, School of Pharmacy and Biochemistry, University of Buenos Aires, Junin 956, C1113AAD Buenos Aires, Argentina. juanitab@ffyb.uba.ar
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 NH(4) (+) and calcium.
Insights
Mitochondrial dysfunction and apoptosis occur in the hippocampus in a minimal hepatic encephalopathy (MHE) model. This study identifies hippocampal mitochondria as key mediators of apoptosis in MHE.
Area of Science:
- Neuroscience
- Hepatology
- Cell Biology
Background:
- Minimal hepatic encephalopathy (MHE) is associated with neurological complications.
- The role of hippocampal apoptosis and mitochondrial dysfunction in MHE remains unclear.
Purpose of the Study:
- To investigate apoptosis and mitochondrial dysfunction in the hippocampus of an experimental MHE model.
- To elucidate the molecular mechanisms underlying hippocampal apoptosis in MHE.
Main Methods:
- An experimental MHE model was established via portal vein stricture, inducing portal hypertension and hyperammonemia.
- Hippocampal apoptosis was assessed using TUNEL staining, Western blotting for Bcl-2 family proteins, and cytochrome c release assays.
- Mitochondrial function was evaluated through respiration assays, cytochrome c oxidase activity measurements, membrane potential determination, and swelling sensitivity tests.
Main Results:
- MHE animals exhibited increased DNA fragmentation (2.3-fold TUNEL-positive cells) and elevated Bax/Bcl-xL ratios, indicating apoptosis.
- Hippocampal mitochondria showed impaired respiratory control (45% decrease), reduced complex IV activity (46% decrease), and diminished membrane potential (13% decrease).
- Mitochondria from MHE animals were more susceptible to calcium-induced swelling, and in vitro studies suggested ammonia and calcium can trigger mitochondrial permeability transition (MPT) and apoptosis.
Conclusions:
- The study demonstrates apoptosis and mitochondrial dysfunction in the hippocampus of an MHE model.
- Hippocampal mitochondria are identified as crucial mediators of apoptosis in MHE.
- The findings suggest intrinsic apoptotic pathways involving mitochondrial dysfunction are central to MHE-associated neurodegeneration.
