Acetaminophen (APAP) overdose is a common cause of drug-induced liver injury (DILI). Ferroptosis has been recently implicated in APAP-induced liver injury (AILI). However, the functional role and underlying mechanisms of mitochondria in APAP-induced ferroptosis are unclear. In this study, the voltage-dependent anion channel (VDAC) oligomerization inhibitor VBIT-12 and ferroptosis inhibitors were injected via tail vein in APAP-injured mice. Targeted metabolomics and untargeted lipidomic analyses were utilized to explore underlying mechanisms of APAP-induced mitochondrial dysfunction and subsequent ferroptosis. As a result, APAP overdose led to characteristic changes generally observed in ferroptosis. The use of ferroptosis inhibitor ferrostatin-1 (or UAMC3203) and iron chelator deferoxamine further confirmed that ferroptosis was responsible for AILI. Mitochondrial dysfunction, which is associated with the tricarboxylic acid cycle and fatty acid β-oxidation suppression, may drive APAP-induced ferroptosis in hepatocytes. APAP overdose induced VDAC1 oligomerization in hepatocytes, and protecting mitochondria via VBIT-12 alleviated APAP-induced ferroptosis. Ceramide and cardiolipin levels were increased via UAMC3203 or VBIT-12 in APAP-induced ferroptosis in hepatocytes. Knockdown of Smpd1 and Taz expression responsible for ceramide and cardiolipin synthesis, respectively, aggravated APAP-induced mitochondrial dysfunction and ferroptosis in hepatocytes, whereas Taz overexpression protected against these processes. By immunohistochemical staining, we found that levels of 4-hydroxynonenal (4-HNE) protein adducts were increased in the liver biopsy samples of patients with DILI compared to that in those of patients with autoimmune liver disease, chronic viral hepatitis B, and non-alcoholic fatty liver disease (NAFLD). In summary, protecting mitochondria via inhibiting VDAC1 oligomerization attenuated hepatocyte ferroptosis by restoring ceramide and cardiolipin content in AILI.

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中文翻译：

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通过抑制VDAC1寡聚化保护线粒体减轻对乙酰氨基酚致急性肝损伤的铁死亡

对乙酰氨基酚 (APAP) 过量是药物性肝损伤 (DILI) 的常见原因。最近，铁死亡与 APAP 诱导的肝损伤（AILI）有关。然而，线粒体在 APAP 诱导的铁死亡中的功能作用和潜在机制尚不清楚。在这项研究中，电压依赖性阴离子通道 (VDAC) 寡聚化抑制剂 VBIT-12 和铁死亡抑制剂通过尾静脉注射到 APAP 损伤的小鼠中。利用靶向代谢组学和非靶向脂质组学分析来探索 APAP 诱导的线粒体功能障碍和随后的铁死亡的潜在机制。因此，APAP 过量导致铁死亡中普遍观察到的特征性变化。使用铁死亡抑制剂 ferrostatin-1（或 UAMC3203）和铁螯合剂去铁胺进一步证实了铁死亡是导致 AILI 的原因。与三羧酸循环和脂肪酸β-氧化抑制相关的线粒体功能障碍可能驱动APAP诱导的肝细胞铁死亡。APAP 过量诱导肝细胞中的 VDAC1 寡聚化，通过 VBIT-12 保护线粒体可减轻 APAP 诱导的铁死亡。在 APAP 诱导的肝细胞铁死亡中，神经酰胺和心磷脂水平通过 UAMC3203 或 VBIT-12 增加。击倒 通过 VBIT-12 保护线粒体可减轻 APAP 诱导的铁死亡。在 APAP 诱导的肝细胞铁死亡中，神经酰胺和心磷脂水平通过 UAMC3203 或 VBIT-12 增加。击倒 通过 VBIT-12 保护线粒体可减轻 APAP 诱导的铁死亡。在 APAP 诱导的肝细胞铁死亡中，神经酰胺和心磷脂水平通过 UAMC3203 或 VBIT-12 增加。击倒分别负责神经酰胺和心磷脂合成的Smpd1和Taz表达加重了 APAP 诱导的肝细胞线粒体功能障碍和铁死亡，而Taz过表达可防止这些过程。通过免疫组织化学染色，我们发现 DILI 患者的肝活检样本中 4-羟基壬烯醛 (4-HNE) 蛋白加合物的水平高于自身免疫性肝病、慢性病毒性乙型肝炎和非乙型肝炎患者的肝活检样本。酒精性脂肪肝（NAFLD）。总之，通过抑制 VDAC1 寡聚化来保护线粒体可通过恢复 AILI 中的神经酰胺和心磷脂含量来减轻肝细胞铁死亡。

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