Supernatant from Hepatocyte Cultures with Drugs That Cause Idiosyncratic Liver Injury Activates Macrophage Inflammasomes
Supernatant from Hepatocyte Cultures with Drugs That Cause Idiosyncratic Liver Injury Activates Macrophage Inflammasomes
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DOI：
10.1021/acs.chemrestox.7b00065
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发表时间：
2017-06-01
影响因子：
4.1
通讯作者：
Uetrecht, Jack
中科院分区：
文献类型：
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作者：
Kato, Ryuji;Uetrecht, Jack
越来越多的证据表明，大多数特异质药物性肝损伤 (IDILI) 是免疫介导的，并且在大多数情况下，反应性代谢产物似乎是诱导这种免疫反应的原因。反应性代谢物可通过释放损伤相关分子模式 (DAMN) 导致细胞损伤，这被认为与免疫激活有关。据推测，肝脏之所以成为特殊药物反应的常见目标，是因为。它是药物代谢和反应性代谢物形成的主要场所。 DAMPS 可以激活炎症小体，这可能是 DAMPs 启动免疫反应的常见机制。在这项研究中，我们测试了药物诱导激活炎症小体的 DAMP 释放的能力。测试的药物是阿莫地喹和奈韦拉平；两者均与严重 IDILI 的发生率显着相关。肝细胞是人肝癌功能性肝细胞4（FLC-4）细胞系。为了检测炎症小体活化，我们使用了人类巨噬细胞系 THP-1 细胞。我们发现两种药物与 FLC-4 细胞孵育 7 天的上清液导致 THP-1 细胞 caspase-1 活性增加和 IL-1 beta 产生增加。然而，单独使用阿莫地喹也能直接激活THP-1细胞。这可能是因为 THP-1 细胞中的髓过氧化物酶可以将阿米喹啉生物活化为反应性代谢物。相比之下，奈韦拉平需要细胞色素 P450 来形成反应性代谢物，因此需要与肝细胞一起孵育。这些结果支持这样的假设：药物的反应性代谢物可以导致 DAMP 的释放，进而激活炎症小体。炎症小体激活可能是药物激活免疫系统的重要步骤，在某些患者中可能导致 IDILI。我们的体外模型简单方便，可用于评估炎症小体激活，这可能是筛选药物IDILI风险的方法，
There is increasing evidence that most idiosyncratic drug-induced liver injury (IDILI) is immune mediated, and in most cases, reactive-metabolites appear to be responsible for the induction of this immune response. Reactive metabolites can cause cell damage with the release of damage-associated molecular patterns (DAMN), which is thought to be involved in immune activation. Presumably, the reason that the liver is a common target of idiosyncratic drug reactions is because. it is the major site of drug metabolism and reactive metabolite formation. Inflammasomes can be activated by DAMPS, and this may be a common-mechanism by which DAMPs initiate an immune response. In this study, we tested the ability of drugs to induce the release of DAMPs that activate inflammasomes The drugs tested were amodiaquine and nevirapine; both are associated with significant incidences of Severe IDILI. The hepatocytes were a human hepatocarcinoma functional liver,cell-4 (FLC-4) cell line. For the detection of inflammasome activation, we used the human macrophage cell line, THP-1 cells.-We found that the supernatant from the incubation of both drugs with FLC-4 cells for 7 days led to increased caspase-1 activity and production of IL-1 beta by THP-1 cells. However, amodiaquine alone,also directly activated THP-1 cells. This is presumably because the myeloperoxidase in THP-1 cells can bioactivate amediaquine to a reactive metabolite. In contrast, nevirapine requires cytochromes P450 for reactive metabolite formation and therefore required incubation with hepatocytes. These results support the hypothesis that reactive metabolites of drugs can cause the release of DAMPs, which in turn can activate inflammasomes. Inflammasome activation may be an important step in the activation of the immune system by drugs, which in some patients can lead to IDILI. Our in vitro model is simple and convenient for evaluating inflammasome activation, and this may be a method to screen drugs for IDILI risk,