Evaluation of Toxicity of Acetaminophen on Human Hepatocyte Mitochondrial Aerobic Respiration by Cellular Energy Metabolism Analysis

Abstract

Abstract: Objective This study aims to observe the effects of APAP on human hepatocyte mitochondrial aerobic respiration utilizing Seahorse XFe24 cell energy metabolism analysis system, thus to provide a methodological reference for the study of APAP toxicity prevention and mechanism in an energy metabolism way. Methods Human hepatocytes (HepG2) were treated with 1, 2, 4, 8, and 16 mM APAP for 24 h in vitro. Cell proliferation was detected by MTT assay and cell morphology was observed under an inverted microscope; cell protein was quantified by BCA assay. Experiment method in HepG2 cells was established by Seahorse XFe24 cell energy metabolism analysis system, effects and characteristics of different concentrations of APAP on human hepatocyte mitochondrial aerobic respiration were detected. Results The best conditions of Seahorse XFe24 cell energy metabolism analysis system for HepG2 were: 2×104 cells per well in 24 wells plate and the concentration of FCCP was 2 μM. APAP (4, 8, 16 mM) which could suppress cell proliferation in MTT results could interrupt mitochondrial aerobic respiration in human hepatocytes. APAP in 4 mM and 8 mM had a tendency to reduce basal respiration and ATP production in human hepatocytes, and 16 mM APAP can significantly inhibit basal respiration, maximal respiration and ATP synthesis. Conclusion High-dose APAP (16 mM) significantly inhibited cell energy metabolism in human hepatocytes live system and showed significant toxicity on live cell mitochondrial aerobic respiration.

Key words: acetaminophen, Seahorse XFe24, mitochondrial aerobic respiration, hepatotoxicity

CLC Number:

R992

LI Mei, LIU Ning, ZHANG Fan, SUN Hua. Evaluation of Toxicity of Acetaminophen on Human Hepatocyte Mitochondrial Aerobic Respiration by Cellular Energy Metabolism Analysis[J]. Chinese Journal of Pharmacovigilance, 2019, 16(4): 193-198.

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