Abstract：Acetaminophen （APAP）， a widely used non-steroidal anti-inflammatory drug frequently induces hepatotoxicity in clinical practice with limited therapeutic options. Hydroxytyrosol （HT）， a natural phenolic compound， demonstrates significant potential as a therapeutic agent for drug-induced liver injury. In this study， APAP-induced AML-12 cell injury and mouse liver injury models were used to explore the protective effect and mechanism of hydroxytyrosol on APAP-induced liver injury by detecting biomarkers closely related to apoptosis and ferroptosis， pathological sections， changes in targets related to Nrf2/GPX4 and LKB1/AMPK pathways， and molecular mimetic docking of target proteins of HT and Nrf2/GPX4 pathways. The results showed that HT could significantly attenuate APAP-induced liver injury in mice， achieving comparable efficacy to the standard hepatoprotective agent silymarin. HT was able to alleviate the abnormal changes of oxidative stress-related markers， inhibit apoptosis and ferroptosis induced by APAP in AML-12 cells and mouse liver， promote the phosphorylation of LKB1 and AMPK in mouse liver and AML-12 cells， upregulate Nrf2， and promote the expression of GPX4 downstream in mouse liver. These indicated that HT plays a key role in APAP-induced liver injury protection by regulating antioxidant stress response and ferroptosis through the cellular Nrf2/GPX4 signaling pathway.
2025, Vol. 59 
