Acetaminophen-induced hepatotoxicity: different mechanisms of acetaminophen-induced ferroptosis and mitochondrial damage
Acetaminophen-induced hepatotoxicity: different mechanisms of acetaminophen-induced ferroptosis and mitochondrial damage
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对乙酰氨基酚诱导的肝毒性：对乙酰氨基酚诱导的铁死亡和线粒体损伤的不同机制
DOI：
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发表时间：
2020
影响因子：
6.1
通讯作者：
Masafumi Takahashi
中科院分区：
文献类型：
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作者：
Naoya Yamada;Takanori Komada;N. Ohno;Masafumi Takahashi
越来越多的证据表明，铁凋亡是一种新型的受调控细胞死亡形式，与多种病理生理过程有关，包括神经退行性疾病、缺血再灌注损伤和癌症（Conrad and Pratt 2019; Stockwell et al. 2017）。我们最近发现由n-6多不饱和脂肪酸驱动的铁下垂介导对乙酰氨基酚（APAP）诱导的急性肝衰竭（Yamada et al. 2020a）。APAP可诱导肝毒性、脂质过氧化和谷胱甘肽（GSH）耗竭，而这些均可被铁凋亡特异性抑制剂铁抑素-1 （fer1）、铁螯合剂去铁胺和α-生育酚（维生素E）显著抑制，表明肝细胞铁凋亡有助于APAP诱导的肝毒性的发展。已知当谷胱甘肽过氧化物酶4 （GPX4）被谷胱甘肽耗竭直接或间接抑制时，膜磷脂的铁依赖性脂质过氧化会驱动铁下沉（Dixon等人，2012；Stockwell等人，2017）；然而，在铁下垂过程中，细胞膜或细胞器的哪一层负责脂质过氧化仍然存在争议。先前的研究表明，铁下垂过程中的脂质过氧化可发生在质膜、内质网（ER）、线粒体、溶酶体和高尔基体中（Bersuker et al. 2019; Doll et al. 2019; Feng and Stockwell 2018; Gao et al. 2019; Gaschler et al. 2018; Magtanong et al. 2018）。其中，虽然已经证明即使在缺乏线粒体的细胞中也会诱导铁下垂（Gaschler et al. 2018），但Gao等人证明，线粒体损伤在半胱氨酸衍生诱导的铁下垂（即GSH耗损）中很重要，但在直接GPX4抑制诱导的铁下垂中并不重要（Gao et al. 2019）。由于APAP被CYP2E1代谢并产生n -乙酰基-对苯醌亚胺（NAPQI）， NAPQI与谷胱甘肽结合并导致谷胱甘肽耗竭（Jaeschke 2015），线粒体可能参与了APAP诱导的铁凋亡。此外，人们普遍认为NAPQI与线粒体蛋白相互作用并产生其加合物，这被认为是肝毒性的关键（Jaeschke 2015）。为了探讨线粒体在APAP诱导的铁下垂和肝毒性中的作用，我们使用透射电镜（TEM）分析了APAP加或不加fer1处理小鼠肝细胞线粒体的形态学变化。动物实验和透射电镜的详细方案已在前面描述（Yamada et al. 2020a, b）。简单地说，将过夜禁食的C57BL6/J小鼠腹腔注射APAP (200 mg/kg)，并在APAP注射前1小时处理铁下沉特异性抑制剂Fer-1或对照物。在该小鼠模型中，APAP注射后3小时血清AST和ALT升高达到峰值，肝脏3区（小叶中心区）出现严重的组织学损伤，而fe -1处理几乎完全阻止了APAP诱导的血清AST和ALT升高。TEM显示，APAP注射后3小时小鼠肝细胞出现严重的线粒体损伤（图1）。出乎意料的是，在apap注射了fer1的小鼠肝细胞中，许多线粒体扩大和肿胀。由于在注射APAP后72小时内，fe -1治疗可防止小鼠肝损伤和死亡（Yamada et al. 2020a），我们假设即使线粒体肿胀，肝细胞也能存活。透射电镜的发现提出了一个值得讨论的重要问题。如上所述，NAPQI诱导GSH耗竭，导致脂质过氧化，可能发生在线粒体膜上，随后导致铁下垂。另一方面，napqi诱导的线粒体蛋白加合物* Masafumi Takahashi masafumi2@jichi.ac.jp
Accumulating evidence indicates that ferroptosis, a novel form of regulated cell death, is implicated in a wide variety of pathophysiological processes including neurodegenerative diseases, ischemia–reperfusion injury, and cancer (Conrad and Pratt 2019; Stockwell et al. 2017). We have recently found that ferroptosis driven by n-6 polyunsaturated fatty acids mediates acetaminophen (APAP)-induced acute liver failure (Yamada et al. 2020a). APAP administration induces hepatotoxicity, lipid peroxidation, and glutathione (GSH) depletion, and these were markedly suppressed by the ferroptosis-specific inhibitor Ferrostatin-1 (Fer-1), the iron chelator deferoxamine, and α-tocopherol (vitamin E), indicating that ferroptosis in hepatocytes contributes to the development of APAP-induced hepatotoxicity. Ferroptosis is known to be driven by the iron-dependent lipid peroxidation of membrane phospholipids when glutathione peroxidase 4 (GPX4) is directly or indirectly inhibited by GSH depletion (Dixon et al. 2012; Stockwell et al. 2017); however, it is still controversial which membrane of cell or organelle is responsible for lipid peroxidation during ferroptosis. Previous studies proposed that lipid peroxidation during ferroptosis can occur in plasma membrane, endoplasmic reticulum (ER), mitochondria, lysosomes, and Golgi complex (Bersuker et al. 2019; Doll et al. 2019; Feng and Stockwell 2018; Gao et al. 2019; Gaschler et al. 2018; Magtanong et al. 2018). Among them, although ferroptosis has been shown to be induced even in the cells lacking mitochondria (Gaschler et al. 2018), Gao et al. demonstrated that mitochondrial damage is important in cysteine-derivationinduced ferroptosis (i.e., GSH depletion), but not in direct GPX4 inhibition-induced ferroptosis (Gao et al. 2019). Because APAP is metabolized by the CYP2E1 and generates N-acetyl-p-benzoquinone imine (NAPQI), which binds to GSH and leads to GSH depletion (Jaeschke 2015), it is likely that mitochondria is involved in APAP-induced ferroptosis. In addition, it is generally accepted that NAPQI interacts with mitochondrial proteins and generates their adducts, which is considered to be critical for hepatotoxicity (Jaeschke 2015). To explore the involvement of mitochondria in APAPinduced ferroptosis and hepatotoxicity, we performed transmission electron microscopy (TEM) to analyze morphological change of mitochondria in hepatocytes of mice treated with APAP with or without Fer-1. Detail protocols for animal experiments and TEM were described previously (Yamada et al. 2020a, b). Briefly, overnight fasted C57BL6/J mice were injected with APAP (200 mg/kg) intraperitoneally, and the ferroptosis specific inhibitor Fer-1 or vehicle was treated 1 h prior to APAP injection. In this mouse model, serum AST and ALT elevation reached a peak at 3 h after APAP injection and severe histological damage was observed in zone 3 (centrilobular area) of the liver, and treatment with Fer-1 almost completely prevented APAP-induced elevation of serum AST and ALT. TEM showed severe mitochondrial damages in hepatocytes of mice at 3 h after APAP injection (Fig. 1). Unexpectedly, many mitochondria were enlarged and swollen in hepatocytes of APAP-injected mice treated with Fer-1. Because Fer-1 treatment prevented liver injury and lethality of mice up to 72 h after APAP injection (Yamada et al. 2020a), we assume that the hepatocytes can survive even after mitochondria are swollen. The TEM findings raise an important issue to be discussed. As mentioned above, NAPQI induces GSH depletion, leading to lipid peroxidation, presumably in the mitochondrial membrane, and subsequent ferroptosis. On the other hand, NAPQI-induced mitochondrial protein adducts * Masafumi Takahashi masafumi2@jichi.ac.jp
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线粒体在铁死亡中的作用
影响因子：
16
作者：
Gao, Minghui;Yi, Junmei;Jiang, Xuejun
通讯作者：
Jiang, Xuejun
影响因子：
4
作者：
Gaschler MM;Hu F;Feng H;Linkermann A;Min W;Stockwell BR
通讯作者：
Stockwell BR
影响因子：
8.6
作者：
Magtanong, Leslie;Ko, Pin-Joe;Dixon, Scott J.
通讯作者：
Dixon, Scott J.
影响因子：
9.8
作者：
Feng H;Stockwell BR
通讯作者：
Stockwell BR