Tolosa Laia, Gómez-Lechón M José, Jiménez Nuria, Hervás David, Jover Ramiro, Donato M Teresa
Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria La Fe, Valencia 46026, Spain.
Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria La Fe, Valencia 46026, Spain; CIBERehd, FIS, Barcelona 08036, Spain.
Toxicol Appl Pharmacol. 2016 Jul 1;302:1-9. doi: 10.1016/j.taap.2016.04.007. Epub 2016 Apr 16.
Only a few in vitro assays have been proposed to evaluate the steatotic potential of new drugs. The present study examines the utility of HepaRG cells as a cell-based assay system for screening drug-induced liver steatosis. A high-content screening assay was run to evaluate multiple toxicity-related cell parameters in HepaRG cells exposed to 28 compounds, including drugs reported to cause steatosis through different mechanisms and non-steatotic compounds. Lipid content was the most sensitive parameter for all the steatotic drugs, whereas no effects on lipid levels were produced by non-steatotic compounds. Apart from fat accumulation, increased ROS production and altered mitochondrial membrane potential were also found in the cells exposed to steatotic drugs, which indicates that all these cellular events contributed to drug-induced hepatotoxicity. These findings are of clinical relevance as most effects were observed at drug concentrations under 100-fold of the therapeutic peak plasmatic concentration. HepaRG cells showed increased lipid overaccumulation vs. HepG2 cells, which suggests greater sensitivity to drug-induced steatosis. An altered expression profile of transcription factors and the genes that code key proteins in lipid metabolism was also found in the cells exposed to drugs capable of inducing liver steatosis. Our results generally indicate the value of HepaRG cells for assessing the risk of liver damage associated with steatogenic compounds and for investigating the molecular mechanisms involved in drug-induced steatosis.
仅提出了少数体外试验来评估新药的脂肪变性潜力。本研究考察了HepaRG细胞作为一种基于细胞的检测系统用于筛选药物性肝脂肪变性的效用。进行了一项高内涵筛选试验，以评估暴露于28种化合物的HepaRG细胞中的多个毒性相关细胞参数，这些化合物包括据报道通过不同机制导致脂肪变性的药物和非脂肪变性化合物。脂质含量是所有致脂肪变性药物最敏感的参数，而非脂肪变性化合物对脂质水平无影响。除了脂肪蓄积外，在暴露于致脂肪变性药物的细胞中还发现活性氧生成增加和线粒体膜电位改变，这表明所有这些细胞事件都促成了药物性肝毒性。这些发现具有临床相关性，因为大多数效应是在药物浓度低于治疗峰值血浆浓度100倍时观察到的。与HepG2细胞相比，HepaRG细胞显示出脂质过度蓄积增加，这表明其对药物性脂肪变性更敏感。在暴露于能够诱导肝脂肪变性的药物的细胞中还发现了转录因子和编码脂质代谢关键蛋白的基因的表达谱改变。我们的结果总体上表明了HepaRG细胞在评估与致脂肪变性化合物相关的肝损伤风险以及研究药物性脂肪变性所涉及的分子机制方面的价值。
