Acetaminophen (APAP) overdose causes hepatotoxicity involving mitochondrial dysfunction. Previous studies showed that translocation of Fe2+ from lysosomes into mitochondria by the mitochondrial Ca2+ uniporter (MCU) promotes the mitochondrial permeability transition (MPT) after APAP. Here, our Aim was to assess protection by iron chelation and MCU inhibition against APAP hepatotoxicity in mice. C57BL/6 mice and hepatocytes were administered toxic doses of APAP with and without starch-desferal (an iron chelator), minocycline (MCU inhibitor), or N-acetylcysteine (NAC). In mice, starch-desferal and minocycline pretreatment decreased ALT and liver necrosis after APAP by >60%. At 24 h after APAP, loss of fluorescence of mitochondrial rhodamine 123 occurred in pericentral hepatocytes often accompanied by propidium iodide labeling, indicating mitochondrial depolarization and cell death. Starch-desferal and minocycline pretreatment decreased mitochondrial depolarization and cell death by more than half. In cultured hepatocytes, cell killing at 10 h after APAP decreased from 83% to 49%, 35% and 27%, respectively, by 1 h posttreatment with minocycline, NAC, and minocycline plus NAC. With 4 h posttreatment in vivo, minocycline and minocycline plus NAC decreased ALT and necrosis by ~20% and ~50%, respectively, but NAC alone was not effective. In conclusion, minocycline and starch-desferal decrease mitochondrial dysfunction and severe liver injury after APAP overdose, suggesting that the MPT is likely triggered by iron uptake into mitochondria through MCU. In vivo, minocycline and minocycline plus NAC posttreatment after APAP protect at later time points than NAC alone, indicating that minocycline has a longer window of efficacy than NAC.

中文翻译：

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在小鼠体内对乙酰氨基酚过量使用后，抑制铁从溶酶体迁移到线粒体可减轻肝脏损伤：米诺环素的保护作用。

对乙酰氨基酚（APAP）过量会引起肝毒性，涉及线粒体功能障碍。先前的研究表明，线粒体Ca2 +单向转运体（MCU）将Fe2 +从溶酶体转移到线粒体中可促进APAP后线粒体通透性转变（MPT）。在这里，我们的目的是评估铁螯合和针对小鼠APAP肝毒性的MCU抑制作用的保护作用。给C57BL / 6小鼠和肝细胞中毒剂量的APAP，含或不含淀粉延迟剂（铁螯合剂），米诺环素（MCU抑制剂）或N-乙酰半胱氨酸（NAC）。在小鼠中，淀粉延缓和米诺环素预处理可使APAP后的ALT和肝坏死减少60％以上。APAP后24小时，线粒体若丹明123的荧光丧失发生在中心周肝细胞中，并经常伴有碘化丙啶标记，表明线粒体去极化和细胞死亡。淀粉延缓和米诺环素预处理可使线粒体去极化和细胞死亡减少一半以上。在培养的肝细胞中，用米诺环素，NAC和米诺环素加NAC处理1小时后，APAP后10小时的细胞杀伤率分别从83％降至49％，35％和27％。体内治疗4小时后，米诺环素和米诺环素加NAC分别使ALT和坏死降低了约20％和〜50％，但单独使用NAC无效。总之，米诺环素和延缓淀粉降解可减轻APAP过量后的线粒体功能障碍和严重的肝损伤，这表明MPT可能是由铁通过MCU吸收到线粒体中引起的。在体内，APAP后的米诺环素和米诺环素加NAC后处理比单独使用NAC的保护时间要晚一些，