PDF(Sci-hub)
Abstract：
Idiosyncratic drug-induced liver injury is a clinical concern with serious consequences. Although many preclinical screening methods have been proposed, it remains difficult to identify compounds associated with this rare but potentially fatal liver condition. Here, we propose a novel assay system to assess the risk of liver injury. Rat primary hepatocytes were cultured in a sandwich configuration, which enables the formation of a typical bile canalicular network. From day 2 to 3, test drugs, mostly selected from a list of cholestatic drugs, were administered, and the length of the network was semi-quantitatively measured by immunofluorescence. Liver injury risk information was collected from drug labels and was compared with in vitro measurements. Of 23 test drugs examined, 15 exhibited potent inhibition of bile canalicular network formation (<60% of control). Effects on cell viability were negligible or minimal as confirmed by lactate dehydrogenase leakage and cellular ATP content assays. For the potent 15 drugs, IC50 values were determined. Finally, maximum daily dose divided by the inhibition constant gave good separation of the highest risk of severe liver toxicity drugs such as troglitazone, benzbromarone, flutamide, and amiodarone from lower risk drugs. In conclusion, inhibitory effect on the bile canalicular network formation observed in in vitro sandwich cultured hepatocytes evaluates a new aspect of drug toxicity, particularly associated with aggravation of liver injury.
摘要：
特异性药物性肝损伤是临床关注的一个严重后果。尽管已经提出了许多临床前筛查方法，目前仍难以确定与这种罕见但可能致命的肝脏疾病相关的化合物.这里,我们提出了一种新的检测系统来评估肝损伤的风险。大鼠原代肝细胞以夹心结构培养，能够形成典型的胆管网络。从第2天到第3天，测试药物，主要选自一系列胆汁淤积药物,被管理，并通过免疫荧光半定量测量网络的长度。从药物标签收集肝损伤风险信息，并与体外测量结果进行比较。在检查的23种测试药物中，图15显示出对胆小管网络形成的有效抑制(<60%的对照)。通过乳酸脱氢酶泄漏和细胞ATP含量测定证实，对细胞活力的影响可忽略不计或最小。对于有效的15种药物，测定IC50值。最后,最大日剂量除以抑制常数可以很好地分离出严重肝毒性药物如曲格列酮的最高风险，苯溴马隆,氟他胺,和低风险药物的胺碘酮。总之,在体外夹心培养肝细胞中观察到的对胆管网络形成的抑制作用评估了药物毒性的新方面，特别是与肝损伤加重有关。
