Species-specific toxicity of diclofenac and troglitazone in primary human and rat hepatocytes
Species-specific toxicity of diclofenac and troglitazone in primary human and rat hepatocytes
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DOI：
10.1016/j.cbi.2008.10.031
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发表时间：
2009-04-15
影响因子：
5.1
通讯作者：
Mueller, Stefan O.
中科院分区：
文献类型：
--
作者：
Lauer, Birthe;Tuschl, Gregor;Mueller, Stefan O.
曲格列酮在被批准用于II型糖尿病治疗后不久就退出了市场，因为它对人有强烈的肝毒性作用，而这种作用无法从动物或体外研究中预测。另一种经常与肝脏不良反应相关的药物，有时会导致急性肝衰竭，是广泛使用的非甾体抗炎药（NSAID）双氯芬酸。由于潜在的分子机制尚不完全清楚，我们用不同剂量的曲格列酮和双氯芬酸处理原代大鼠和人肝细胞单层培养物24小时，以分析与体外毒性相关的物种差异。无严重不良反应的降糖药二甲双胍作为阴性对照。人肝细胞对曲格列酮的敏感性高于大鼠肝细胞，而双氯芬酸在相当相似的浓度下诱导细胞毒性。通过与细胞色素P450 (CYP) 2C和CYP3A特异性抑制剂（参与肝脏外源代谢的主要I相酶）共同处理，我们可以证实CYP3A在曲格列酮生物活化中的突出作用，以及CYP3A和CYP2C在双氯芬酸活化中的作用。抑制这些酶增加了两个物种处理细胞的活力。此外，我们能够证明曲格列酮处理的大鼠和人肝细胞的基因表达模式存在显著的物种差异。与大鼠肝细胞相比，人细胞显示出多种CYPs、外源代谢调节因子和氧化应激标记基因的明显上调。相比之下，双氯芬酸处理的大鼠和人肝细胞的基因表达变化非常相似。总之，我们的研究表明，通过使用来自不同物种的原代肝细胞培养物，结合基因表达谱，可以解决物种特异性效应以及化合物作用模式的适应症。2008爱思唯尔爱尔兰有限公司版权所有。
Troglitazone was withdrawn from the market shortly after approval for diabetes type II therapy because of strong hepatotoxic effects in man that could not be predicted from regulatory animal or in vitro studies. Another pharmaceutical that is regularly associated with adverse effects on the liver, sometimes leading to acute liver failure, is the widely used non-steroidal anti-inflammatory drug (NSAID) diclofenac. Since the underlying molecular mechanisms are not yet fully known, we treated primary rat and human hepatocyte monolayer cultures for 24 h with different doses of troglitazone and diclofenac to analyze species differences related to toxicity in vitro. Metformin an antidiabetic drug which does not cause severe adverse reactions served as negative control. Human hepatocytes showed a higher sensitivity to troglitazone than rat hepatocytes, while diclofenac-induced cytotoxicity at fairly similar concentrations. By co-treatment with specific inhibitors for cytochrome P450 (CYP) 2C and CYP3A - the major phase I enzymes involved in liver xenobiotic metabolism - we could confirm the prominent role of CYP3A in the bioactivation of troglitazone as well as the role of CYP3A and CYP2C in the activation of diclofenac. Inhibition of these enzymes increased the viability of treated cells in both species. Furthermore, we were able to demonstrate marked species differences in gene expression patterns of troglitazone treated rat and human hepatocytes. In contrast to rat hepatocytes, human cells showed distinct upregulation of various CYPs, regulators of xenobiotic metabolism and marker genes for oxidative stress. In contrast, gene expression alterations in rat and human hepatocytes treated with Diclofenac were rather similar. Altogether our study showed that species-specific effects as well as indications for the mode of action of compounds can be addressed by the use of primary hepatocyte cultures from various species in combination with gene expression profiling. (C) 2008 Elsevier Ireland Ltd. All rights reserved.