Unidad de Hepatología Experimental, Torre A, Instituto Investigación Sanitaria La Fe (IIS La Fe), Bulevard Sur s/n, 46026, Valencia, Spain.
Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Valencia, 46010, Valencia, Spain.
Arch Toxicol. 2015 Oct;89(10):1847-60. doi: 10.1007/s00204-014-1334-3. Epub 2014 Aug 27.
A frequent mechanism for drug-induced liver injury (DILI) is mitochondrial impairment, and early evaluation of new drugs for their potential to cause mitochondrial dysfunction is becoming an important task for drug development. To this end, we designed a high-content screening assay to study mitochondrial-induced hepatotoxicity in HepG2 cells in detail. Simultaneous assessment of mitochondrial mass and cell viability in cells exposed for 24 h to compounds provides preliminary information on the mitochondrial- or nonmitochondrial-related hepatotoxic potential of compounds. To fully address the mechanisms implicated in mitochondrial impairment, prelethal changes in mitochondrial superoxide production, mitochondrial membrane potential, mitochondrial permeability transition, intracellular calcium concentration and apoptotic cell death were studied in cells incubated for 1 h with compounds. The assay correctly classified a set of well-known mitochondrial toxicants and negative controls and revealed high sensitivity for the detection of mitochondrial DILI and the establishment of different mitochondrial toxicity risks (low to high). This procedure was used for analysing the potential mitochondrial impairment of six statins to determine their clinical risk. All the tested statins produced mitochondrial impairment, although they showed different levels of toxicity (low-medium toxicity risk). The results suggest that this cell-based assay is a promising in vitro approach to predict the potential of drug candidates to induce mitochondrial-associated hepatotoxicity.
药物性肝损伤（DILI）的一个常见机制是线粒体损伤，因此早期评估新药是否有可能引起线粒体功能障碍，已成为药物开发的一项重要任务。为此，我们设计了一种高通量筛选测定法，以详细研究 HepG2 细胞中的线粒体诱导性肝毒性。在化合物作用 24 小时后，同时评估线粒体质量和细胞活力，可初步了解化合物的线粒体或非线粒体相关肝毒性潜力。为了全面阐明与线粒体损伤相关的机制，在化合物孵育 1 小时后，研究了线粒体中超氧化物产生、线粒体膜电位、线粒体通透性转换、细胞内钙离子浓度和凋亡性细胞死亡的预致死变化。该测定法正确分类了一组已知的线粒体毒性剂和阴性对照物，并显示出对检测线粒体 DILI 和建立不同线粒体毒性风险（低至高）的高灵敏度。该方法用于分析六种他汀类药物的潜在线粒体损伤，以确定其临床风险。所有测试的他汀类药物都产生了线粒体损伤，尽管它们表现出不同程度的毒性（低至中度毒性风险）。结果表明，这种基于细胞的测定法是一种很有前途的体外方法，可用于预测候选药物诱导与线粒体相关的肝毒性的潜力。
