Hadi Mackenzie, Westra Inge M, Starokozhko Viktoriia, Dragovic Sanja, Merema Marjolijn T, Groothuis Geny M M
Division of Pharmacokinetics, Toxicology and Targeting, Department of Pharmacy, University of Groningen , Groningen, The Netherlands.
Chem Res Toxicol. 2013 May 20;26(5):710-20. doi: 10.1021/tx300519p. Epub 2013 Apr 19.
Idiosyncratic drug-induced liver injury (IDILI) is a major problem during drug development and has caused drug withdrawal and black-box warnings. Because of the low concordance of the hepatotoxicity of drugs in animals and humans, robust screening methods using human tissue are needed to predict IDILI in humans. According to the inflammatory stress hypothesis, the effects of inflammation interact with the effects of a drug or its reactive metabolite, precipitating toxic reactions in the liver. As a follow-up to our recently published mouse precision-cut liver slices model, an ex vivo model involving human precision-cut liver slices (hPCLS), co-incubated for 24 h with IDILI-related drugs and lipopolysaccharide (LPS), was developed to study IDILI mechanisms related to inflammatory stress in humans and to detect potential biomarkers. LPS exacerbated the effects of ketoconazole and clozapine toxicity but not those of their non-IDILI-related comparators, voriconazole and olanzapine. However, the IDILI-related drugs diclofenac, carbamazepine, and troglitazone did not show synergistic toxicity with LPS after incubation for 24 h. Co-incubation of ketoconazole and clozapine with LPS decreased the levels of glutathione in hPCLS, but this was not seen for the other drugs. All drugs affected LPS-induced cytokine release, but interestingly, only ketoconazole and clozapine increased the level of LPS-induced TNF release. Decreased levels of glutathione and cysteine conjugates of clozapine were detected in IDILI-responding livers following cotreatment with LPS. In conclusion, we identified ketoconazole and clozapine as drugs that exhibited synergistic toxicity with LPS, while glutathione and TNF were found to be potential biomarkers for IDILI-inducing drugs mediated by inflammatory stress. hPCLS appear to be suitable for further unraveling the mechanisms of inflammatory stress-associated IDILI.
特异质性药物性肝损伤(IDILI)是药物研发过程中的一个主要问题,已导致药物撤市和黑框警告。由于药物在动物和人类中的肝毒性一致性较低,因此需要使用人体组织的可靠筛选方法来预测人类的IDILI。根据炎症应激假说,炎症作用与药物或其反应性代谢产物的作用相互影响,从而在肝脏中引发毒性反应。作为我们最近发表的小鼠精密肝切片模型的后续研究,我们开发了一种涉及人精密肝切片(hPCLS)的体外模型,该模型与IDILI相关药物和脂多糖(LPS)共同孵育24小时,以研究与人类炎症应激相关的IDILI机制并检测潜在的生物标志物。LPS加剧了酮康唑和氯氮平的毒性作用,但未加剧其非IDILI相关对照药伏立康唑和奥氮平的毒性作用。然而,IDILI相关药物双氯芬酸、卡马西平和曲格列酮在孵育24小时后未显示出与LPS的协同毒性。酮康唑和氯氮平与LPS共同孵育降低了hPCLS中的谷胱甘肽水平,但其他药物未出现这种情况。所有药物均影响LPS诱导的细胞因子释放,但有趣的是,只有酮康唑和氯氮平增加了LPS诱导的TNF释放水平。在与LPS联合治疗后的IDILI反应性肝脏中检测到氯氮平的谷胱甘肽和半胱氨酸共轭物水平降低。总之,我们确定酮康唑和氯氮平为与LPS表现出协同毒性的药物,而谷胱甘肽和TNF被发现是由炎症应激介导的IDILI诱导药物的潜在生物标志物。hPCLS似乎适合进一步阐明炎症应激相关IDILI的机制。