To investigate the protective effect of ligustrazine on the liver injury induced by acetaminophen(APAP) and the underlying mechanism.Methods:Sixty specific pathogen free(SPF)-grade male C57BL/6 mice of 7 to 8 weeks old were randomized into control，model，low-，medium-，high-dose(30，100，and 300 mg/kg，respectively) ligustrazine，and positive control(150 mg/kg N-acetylcysteine) group，with 10 mice in each group.Mice were administrated with corresponding drugs by gavage for 12 consecutive days.Two hours after the last administration，mice in other groups except the control group were intraperitoneally injected with 300 mg/kg APAP for modeling，and samples were collected 14 h later.Results:Compared with the control group，the model group showed increased liver index，elevated levels of iron ion，malondialdehyde(MDA)，acyl-CoA synthetase long-chain family member 4(ACSL4)，pro-inflammatory cytokines，and chemokines(all P<0.05)，and down-regulated protein levels of nuclear E2-related factor 2(Nrf2)，NAD(P)H:quinone oxidoreductase 1(NQO1)，heme oxygenase-1(HO-1)，and glutathione peroxidase(GPX4)(all P<0.05).Compared with the model group，ligustrazine decreased the liver index，lowered the levels of iron ion，MDA，ACSL4，pro-inflammatory cytokines，and chemokines(all P<0.05)，and up-regulated the protein levels of Nrf2，NQO1，HO-1，and GPX4(all P<0.05).Conclusion:Ligustrazine can alleviate the APAP-induced liver injury by inhibiting inflammatory response and ferroptosis pathway，which may be associated with the activation of the Nrf2/NQO1/HO-1 pathway.