Mitochondrial dysfunction induced by leflunomide and its active metabolite.
Mitochondrial dysfunction induced by leflunomide and its active metabolite.
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DOI：
10.1016/j.tox.2018.02.003
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发表时间：
2018-03-01
期刊：
影响因子：
4.5
通讯作者：
Guo L
中科院分区：
文献类型：
--
作者：
Xuan J;Ren Z;Qing T;Couch L;Shi L;Tolleson WH;Guo L
来氟米特是一种用于治疗类风湿性关节炎的抗炎药，已经被标记了一个黑框警告，警告它会增加肝损伤的风险。来氟米特的活性代谢产物A771726也带有关于潜在肝毒性的黑框警告，已作为特立氟胺上市，用于治疗复发性多发性硬化症。然而，到目前为止，与这两种药物相关的肝损伤机制仍然难以捉摸。在这项研究中，细胞毒性试验表明，ATP消耗和随后的LDH释放诱导的时间和浓度依赖性的方式来氟米特在HepG2细胞，并在较小程度上，A77 1726。当半乳糖取代葡萄糖作为糖源时，来氟米特引起的细胞ATP水平的下降急剧加剧，表明来氟米特潜在的线粒体责任。通过测量免疫捕获的线粒体氧化磷酸化（OXPHOS）复合物的活性，我们发现来氟米特和A77 1726优先靶向复合物V（F1FO ATP合成酶），IC 50值分别为35.0和63.7 μM。Bongkrekic酸是一种靶向腺嘌呤核苷酸移位酶的线粒体通透性转换孔阻滞剂，可显著减弱来氟米特和A77 1726诱导的线粒体膜去极化、ATP耗竭和LDH渗漏。在来氟米特处理的HepG2细胞中观察到线粒体功能在转录水平上的实质性改变，而A77 1726对细胞转录组的影响则不那么深刻。我们的研究结果表明，线粒体功能障碍可能与来氟米特和A77 1726相关的肝毒性有关，前者表现出更高的毒性效力。
Leflunomide, an anti-inflammatory drug used for the treatment of rheumatoid arthritis, has been marked with a black box warning regarding an increased risk of liver injury. The active metabolite of leflunomide, A771726, which also carries a boxed warning about potential hepatotoxicity, has been marketed as teriflunomide for the treatment of relapsing multiple sclerosis. Thus far, however, the mechanism of liver injury associated with the two drugs has remained elusive. In this study, cytotoxicity assays showed that ATP depletion and subsequent LDH release were induced in a time- and concentration-dependent manner by leflunomide in HepG2 cells, and to a lesser extent, by A77 1726. The decline of cellular ATP levels caused by leflunomide was dramatically exacerbated when galactose was substituted for glucose as the sugar source, indicating a potential mitochondrial liability of leflunomide. By measuring the activities of immuno-captured mitochondrial oxidative phosphorylation (OXPHOS) complexes, we found that leflunomide and A77 1726 preferentially targeted complex V (F1FO ATP synthase), with IC50 values of 35.0 and 63.7 μM, respectively. Bongkrekic acid, a mitochondrial permeability transition pore blocker that targets adenine nucleotide translocase, profoundly attenuated mitochondrial membrane depolarization, ATP depletion, and LDH leakage induced by leflunomide and A77 1726. Substantial alterations of mitochondrial function at the transcript level were observed in leflunomide-treated HepG2 cells, whereas the effects of A77 1726 on the cellular transcriptome were much less profound. Our results suggest that mitochondrial dysfunction may be implicated in the hepatotoxicity associated with leflunomide and A77 1726, with the former exhibiting higher toxicity potency.
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影响因子：
4.3
作者：
Bonora, Massimo;Bononi, Angela;Pinton, Paolo
通讯作者：
Pinton, Paolo
影响因子：
4.8
作者：
Fiermonte, G;De Leonardis, F;Palmieri, F
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作者：
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影响因子：
3.9
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
Baines, Christopher P.