Objective: To investigate the mechanism by which Notch1 signaling regulates APAP-induced liver injury (AILI) via TAK1. Methods: Twenty-four C57BL/6, floxed Notch1（Notch1FL/FL）， and Myeloid-specific Notch1 knockout（Notch1M-KO）mice were randomly divided into four groups: control (Notch1FL/FL) + PBS (n=6), control + APAP (n=6), Notch1M-KO + PBS (n=6), and Notch1M-KO + APAP (n=6), and APAP was injected intraperitoneally to establish the AILI model. Mouse serum and tissue samples were retained for serological examination, HE staining, immunofluorescence, Western blot(WB), and enzyme-linked immunoassay analysis, respectively. Results: Compared with the Notch1FL/FL group, the Notch1M-KO group displayed significantly higher serum levels of ALT, AST, and cytokines; HE staining reveals significant hepatocyte edema, sinusoidal stasis, and increased macrophage accumulation; the DCF probe demonstrated increased intracellular ROS expression. Elevated TAK1 activated p-P65 and increased RIPK1, and p-MLKL expression, thus promoting the development of necroptosis in hepatocytes and aggravating the liver injury. At the same time, caspase-8 expression was suppressed. Conclusion: In AILI, myeloid-specific Notch1 knockout activates TAK1 expression, which also decreases Caspase-8 levels and promotes the RIPK1-MLKL necroptosis pathway, aggravating the liver injury.