Borlak Jürgen, Chougule Anil, Singh Prafull Kumar
Centre for Pharmacology and Toxicology, Hannover Medical School, Carl-Neuberg-Straße 1, 30625 Hannover, Germany.
Toxicol In Vitro. 2014 Aug;28(5):784-95. doi: 10.1016/j.tiv.2014.03.006. Epub 2014 Mar 28.
In preclinical hepatotoxicity testing cell based assays are frequently employed. However, prediction of clinical drug induced liver injury (DILI) remains a major challenge. Here we examined the usefulness of frequently employed markers of hepatocellular injury in cultures of primary human hepatocytes (PHH) in response to treatment with either paracetamol, rifampicin, petadolex and/or amiodarone. The changes in the metabolic competency (urea and albumin) and cellular injury (AST, ALT, ALP, LDH, γGT and succinate dehydrogenase) were determined at therapeutic and above drug concentrations as to evaluate the utility of these markers in in vitro systems. Initially, treatment of PHH with any of the drugs caused a statistically significant reduction in enzyme activities to suggest a switch from basic amino acid metabolism towards induced detoxification. However, treatment for prolonged periods of time caused cytolysis, as evidenced by the significant rise in extracellular LDH and the concomitant increase in ALT and AST activity. Notably, amongst the various endpoints studied, urea was best to demonstrate dose dependent metabolic stress, while other markers of hepatocellular injury were highly variable. Taken collectively, urea measurement proofed to be robust in predicting hepatocellular stress; therefore it should be included in preclinical testing strategies for an improved prediction of DILI.
在临床前肝毒性测试中，基于细胞的检测方法经常被采用。然而，预测临床药物性肝损伤（DILI）仍然是一项重大挑战。在此，我们研究了常用的肝细胞损伤标志物在原代人肝细胞（PHH）培养物中对乙酰氨基酚、利福平、Petadolex和/或胺碘酮处理的反应中的有用性。在治疗浓度及以上药物浓度下测定代谢能力（尿素和白蛋白）和细胞损伤（AST、ALT、ALP、LDH、γGT和琥珀酸脱氢酶）的变化，以评估这些标志物在体外系统中的效用。最初，用任何一种药物处理PHH都会导致酶活性出现统计学上的显著降低，表明从碱性氨基酸代谢转向诱导解毒。然而，长时间处理会导致细胞溶解，细胞外LDH的显著升高以及ALT和AST活性的随之增加证明了这一点。值得注意的是，在研究的各种终点中，尿素最能证明剂量依赖性代谢应激，而其他肝细胞损伤标志物变化很大。总体而言，尿素测量被证明在预测肝细胞应激方面很可靠；因此，应将其纳入临床前测试策略中，以改进对DILI的预测。
