Oxidative stress-dependent frataxin inhibition mediated alcoholic hepatocytotoxicity through ferroptosis
Oxidative stress-dependent frataxin inhibition mediated alcoholic hepatocytotoxicity through ferroptosis
复制标题
氧化应激依赖性 frataxin 抑制通过铁死亡介导酒精性肝细胞毒性
DOI：
10.1016/j.tox.2020.152584
复制
发表时间：
2020-12-01
期刊：
影响因子：
4.5
通讯作者：
Tang, Yuhan
中科院分区：
文献类型：
--
作者：
Liu, Jingjing;He, Hui;Tang, Yuhan
酒精性肝病（ALD）是一种严重的肝脏疾病，发病率和死亡率都很高。然而，主要参与铁稳态和氧化应激的线粒体蛋白frataxin在ALD的发病机制中仍不确定。在本研究中，我们探讨了frataxin在ALD中的作用。乙醇（100 mM）可降低HepG2细胞48和72 h时frataxin的表达。引人注意的是，在细胞色素P450 2E1过表达的HepG2（HepG2(CYP2E1+/+)）中，乙醇刺激12 h后，frataxin水平下调。此外，通过Lieber-DeCarli液体饲料（总热量的30%）长期饲喂乙醇15周，可显著抑制frataxin的表达。凋亡特征蛋白异常，线粒体形态损伤，肝脏丙二醛增强，谷胱甘肽减少，原代肝细胞活性氧和线粒体不稳定铁池积累。值得注意的是，蛋白质组学筛选fraataxin缺陷- hepg2进一步表明fraataxin与铁下垂有关。此外，铁下垂抑制剂铁抑素-1阻断了乙醇在HepG2（CYP2E1+/+）中乳酸脱氢酶释放的增加。最重要的是，通过评估乳酸脱氢酶水平、细胞形态学改变、线粒体不稳定铁池和脂质过氧化，fraataxin缺乏增强了乙醇驱动的铁死亡。相反，恢复frataxin可减轻对铁下垂的敏感性。此外，frataxin过表达减轻了乙醇诱导的HepG2（CYP2E1+/+）铁下垂的敏感性。总的来说，我们的研究揭示了frataxin介导的铁下垂有助于ALD，强调了ALD的潜在治疗策略。
Alcoholic liver disease (ALD) is one of the severe liver diseases, resulting in high morbidity and mortality. However, frataxin, a mitochondrial protein mainly participating in iron homeostasis and oxidative stress, remains uncertain in the pathogenesis of ALD. In the present study, the role of frataxin in ALD was investigated. Ethanol (100 mM) decreased frataxin expression at 48 and 72 h in HepG2. Dramatically, in HepG2 over-expressing cytochrome P450 2E1 (HepG2(CYP2E1+/+)), frataxin level was down-regulated with ethanol stimulation at 12 h. Moreover, chronically feeding ethanol to mice via Lieber-DeCarli liquid diet (30 % of total calories) for 15 weeks significantly inhibited frataxin expression. Ferroptosis signature proteins were dysregulated, accompanied by mitochondrial damage of morphology, enhanced malondialdehyde and decreased glutathione in the liver, as well as accumulation of reactive oxygen species and mitochondrial labile iron pool in primary hepatocytes. Notably, proteomics screening of frataxin deficient-HepG2 further suggested frataxin was associated with ferroptosis. Furthermore, the ferroptosis inhibitor ferrostatin-1 blocked the increase of lactate dehydrogenase release by ethanol in HepG2(CYP2E1+/+). Most importantly, frataxin deficiency enhanced ferroptosis driven by ethanol via evaluating the levels of lactate dehydrogenase, cell morphological changes, mitochondrial labile iron pool, and lipid peroxidation. Conversely, restoring frataxin alleviated the sensitivity to ferroptosis. In addition, frataxin overexpression mitigated the sensitivity of ethanol-induced ferroptosis in HepG2(CYP2E1+/+). Collectively, our study revealed that frataxin-mediated ferroptosis contributed to ALD, highlighting a potential therapeutic strategy for ALD.