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Generation of a Rat Model of Acute Liver Failure by Combining 70% Partial Hepatectomy and Acetaminophen
Published on: November 27, 2019
Comparison of acetaminophen toxicity in primary hepatocytes isolated from transgenic mice with different
V Mezera1, O Kucera2, A Moravcova2
1Department of Physiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic. mezev5ar@lfhk.cuni.cz.
Abstract:
The nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor, important for combating electrophilic and oxidative stress in the liver and other organs. This encompasses detoxification of hepatotoxic drugs, including acetaminophen (APAP). Recently, an association between apolipoprotein E (ApoE) genotype and Nrf2 expression was described. We compared the toxicity of APAP on primary culture hepatocytes isolated from transgenic mice carrying two different human ApoE alleles and wild-type controls. The cells were exposed to APAP in concentrations from 0.5 to 4 mM for up to 24 hours. APAP led to a dose-dependent hepatotoxicity from 1 mM after 16 h exposure in all mice tested. The toxicity was higher in hepatocytes isolated from both transgenic strains than in wild-type controls and most pronounced in ApoE3 mice. Concurrently, there was a decline in mitochondrial membrane potential, especially in ApoE3 hepatocytes. The formation of reactive oxygen species was increased after 24 hours with 2.5 mM APAP in hepatocytes of all strains tested, with the highest increase being in the ApoE3 genotype. The activity of caspases 3 and 7 did not differ among groups and was minimal after 24 hour incubation with 4 mM APAP. We observed higher lipid accumulation in hepatocytes isolated from both transgenic strains than in wild-type controls. The expression of Nrf2-dependent genes was higher in ApoE3 than in ApoE4 hepatocytes and some of these genes were induced by APAP treatment. In conclusion, transgenic mice with ApoE4 and ApoE3 alleles displayed higher susceptibility to acute APAP toxicity in vitro than wild-type mice. Of the two transgenic genotypes tested, ApoE3 allele carriers were more prone to injury.
Insights
Apolipoprotein E (ApoE) genotype influences acetaminophen (APAP) liver toxicity. ApoE3 and ApoE4 mice showed increased APAP toxicity and oxidative stress compared to controls, with ApoE3 being most susceptible.
Area of Science:
- Hepatology
- Toxicology
- Genetics
Background:
- Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates cellular defense against oxidative and electrophilic stress, including drug detoxification.
- Apolipoprotein E (ApoE) genotype has been linked to Nrf2 expression, suggesting a potential role in drug metabolism and toxicity.
Purpose of the Study:
- To investigate the impact of different human apolipoprotein E (ApoE) alleles (ApoE3 and ApoE4) on acetaminophen (APAP)-induced hepatotoxicity in primary mouse hepatocytes.
- To explore the relationship between ApoE genotype, Nrf2-dependent gene expression, oxidative stress, and mitochondrial function during APAP exposure.
Main Methods:
- Primary hepatocytes were isolated from wild-type, ApoE3, and ApoE4 transgenic mice.
- Hepatocytes were exposed to varying concentrations of APAP (0.5–4 mM) for up to 24 hours.
- Assessed APAP toxicity, mitochondrial membrane potential, reactive oxygen species (ROS) formation, caspase activity, lipid accumulation, and Nrf2-dependent gene expression.
Main Results:
- APAP induced dose-dependent hepatotoxicity in all tested genotypes, with significantly higher toxicity observed in ApoE3 and ApoE4 hepatocytes compared to wild-type controls.
- ApoE3 hepatocytes exhibited the most pronounced toxicity, increased ROS formation, and decline in mitochondrial membrane potential.
- Higher lipid accumulation and increased expression of Nrf2-dependent genes were noted in transgenic hepatocytes, with some genes induced by APAP treatment.
Conclusions:
- Transgenic mice carrying human ApoE3 and ApoE4 alleles demonstrate heightened susceptibility to acute APAP-induced hepatotoxicity in vitro.
- The ApoE3 genotype confers a greater proneness to APAP-induced liver injury compared to ApoE4 and wild-type genotypes, potentially mediated by altered oxidative stress and Nrf2 pathway regulation.
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