Toxicological Centre, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium; Research group In Vitro Toxicology and Dermato-Cosmetology (IVTD), Vrije Universiteit Brussel, Laarbeeklaan 103, 1090, Jette, Belgium.
Department of Mathematics & Computer Science, University of Antwerp, Middelheimlaan 1, 2020 Antwerp, Belgium; Biomedical Informatics Network Antwerpen (Biomina), University of Antwerp, Middelheimlaan 1, 2020 Antwerp, Belgium.
Toxicol Appl Pharmacol. 2019 Sep 15;379:114666. doi: 10.1016/j.taap.2019.114666. Epub 2019 Jul 16.
Cholestasis is a liver disease associated with retention of bile in the liver, which leads to local hepatic inflammation and severe liver damage. In order to investigate the mode of action of drug-induced cholestasis, in vitro models have shown to be able to recapitulate important elements of this disease. In this study, we applied untargeted metabolomics to investigate the metabolic perturbances in HepaRG® cells exposed for 24 h and 72 h to bosentan, a cholestatic reference toxicant. Intracellular profiles were extracted and analysed with liquid chromatography and accurate-mass spectrometry. Metabolites of interest were selected using partial least-squares discriminant analysis and random forest classifier models. The observed metabolic patterns associated with cholestasis in vitro were complex. Acute (24 h) exposure revealed metabolites related to apoptosis, such as ceramide and triglyceride accumulation, in combination with phosphatidylethanolamine, choline and carnitine depletion. Metabolomic alterations during exposure to lower dosages and a prolonged exposure (72 h) included carnitine upregulation and changes in the polyamine metabolism. These metabolites were linked to changes in phospholipid metabolism, mitochondrial pathways and energy homeostasis. The metabolic changes confirmed the mitotoxic effects of bosentan and revealed the potential involvement of phospholipid metabolism as part of the mode of action of drug-induced cholestasis.
胆汁淤积症是一种与胆汁在肝脏中滞留有关的肝脏疾病，可导致局部肝炎症和严重的肝损伤。为了研究药物性胆汁淤积症的作用机制，体外模型已被证明能够重现该疾病的重要元素。在这项研究中，我们应用非靶向代谢组学方法来研究 HepaRG®细胞在暴露于波生坦（一种胆汁淤积的参考毒性物质）24 小时和 72 小时后的代谢紊乱。用液相色谱和精确质量质谱法提取和分析细胞内谱。使用偏最小二乘判别分析和随机森林分类器模型选择感兴趣的代谢物。与体外胆汁淤积相关的观察到的代谢模式是复杂的。急性（24 小时）暴露显示与细胞凋亡相关的代谢物，如神经酰胺和甘油三酯积累，同时伴有磷脂酰乙醇胺、胆碱和肉碱耗竭。在较低剂量和延长暴露（72 小时）期间，代谢组学的改变包括肉碱上调和多胺代谢的改变。这些代谢物与磷脂代谢、线粒体途径和能量稳态的变化有关。这些代谢变化证实了波生坦的线粒体毒性作用，并揭示了磷脂代谢可能是药物性胆汁淤积症作用机制的一部分。
