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Acetaminophen (APAP) overdose can cause liver injury in animals and human beings by inducing mitochondrial damage, which need to be further evaluated. The aim of the current study is to assess the changes between oxidative damage mitophagy in vivo and in vitro, which mimics APAP\u2010induced liver injury (AILI) in humans. Liver damage was monitored by measuring the levels of biochemical indexes. Proteins associated with oxidative stress were inspected by western blot analysis. After given APAP to both Nrf2<jats:sup>\u2212\/\u2212<\/jats:sup> and wild\u2010type mice, Nrf2<jats:sup>\u2212\/\u2212<\/jats:sup> mice were highly susceptible to APAP induced liver injury. Actually, rapamycin promoted the process of autophagy, reducing the formation of giant mitochondria and lipid droplets. Both tBHQ and NAC protected hepatic cells, promoting Nrf2 translocation into nucleus and increasing the expression of downstream enzymes and proteins. 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