Establishment of a Drug-Induced, Bile Acid-Dependent Hepatotoxicity Model Using HepaRG Cells
Establishment of a Drug-Induced, Bile Acid-Dependent Hepatotoxicity Model Using HepaRG Cells
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DOI：
10.1016/j.xphs.2016.01.013
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发表时间：
2016-04-01
影响因子：
3.8
通讯作者：
Ito, Kousei
中科院分区：
文献类型：
--
作者：
Susukida, Takeshi;Sekine, Shuichi;Ito, Kousei
胆汁酸（BA）滞留在肝细胞内是胆汁淤积性药物诱导的肝损伤（DILI）的潜在机制。我们先前开发了一种使用体外培养的人肝细胞（SCHHs）的试验，以评估伴随细胞内BA蓄积的药物诱导的肝细胞毒性。然而，由于通常与原代人肝细胞的使用相关的缺点（例如，可用性有限、批间变异性和高成本），我们检查了人肝干细胞系HepaRG是否也适用于我们的检测系统。因此，人BA外排和摄取转运蛋白的mRNA表达水平在HepaRG细胞中低于SCHHs，但高于HepG 2人肝癌细胞。然而，HepaRG细胞和SCHHs对22种选定的药物（包括环孢素A（CsA））显示出相似的毒性反应。CsA（10 μ M）在BA存在下对HepaRG细胞具有细胞毒性，并且还将[H-3]牛磺胆酸的胆汁外排率从38.5%降低至19.2%。因此，HepaRG细胞可用于评价胆汁BA外排抑制引起的BA依赖性药物毒性。无论如何，与SCHH相比，HepaRG细胞的胆汁淤积性DILI风险的预测准确性较差，这表明我们的DILI模型系统需要进一步改进以增加HepaRG细胞作为临床前筛选工具的效用。(C)2016年美国药学协会（R）。爱思唯尔公司出版。All rights reserved.
Bile acid (BA) retention within hepatocytes is an underlying mechanism of cholestatic drug-induced liver injury (DILI). We previously developed an assay using sandwich-cultured human hepatocytes (SCHHs) to evaluate drug-induced hepatocyte toxicity accompanying intracellular BA accumulation. However, due to shortcomings commonly associated with the use of primary human hepatocytes (e.g., limited availability, lot-to-lot variability, and high cost), we examined if the human hepatic stem cell line, HepaRG, might also be applicable to our assay system. Consequently, mRNA expression levels of human BA efflux and uptake transporters were lower in HepaRG cells than in SCHHs but higher than in HepG2 human hepatoma cells. Nevertheless, HepaRG cells and SCHHs showed similar toxicity responses to 22 selected drugs, including cyclosporine A (CsA). CsA (10 mu M) was cytotoxic toward HepaRG cells in the presence of BAs and also reduced the biliary efflux rate of [H-3] taurocholic acid from 38.5% to 19.2%. Therefore, HepaRG cells are useful for the evaluation of BA-dependent drug toxicity caused by biliary BA efflux inhibition. Regardless, the prediction accuracy for cholestatic DILI risk was poor for HepaRG cells versus SCHHs, suggesting that our DILI model system requires further improvements to increase the utility of HepaRG cells as a preclinical screening tool. (C) 2016 American Pharmacists Association(R). Published by Elsevier Inc. All rights reserved.