Acute liver injury (ALI) is a significant global public health issue that can rapidly develop into acute liver failure, seriously endangering the safety of patients. Eugenol has various pharmacological effects such as antioxidant, anti-inflammatory, antibacterial, and neuroprotective properties. Through pathological section observation, eugenol can alleviate the degree of liver damage caused by thioacetamide. Based on metabolomics, a total of 87 metabolites were found to have differences in content between the control group and the thioacetamide group. Compared with the control group, the contents of 42 metabolites had increased and 45 metabolites had decreased in the thioacetamide group. These differential expressed metabolites mainly indicate inflammatory damage, oxidative damage, and abnormal energy metabolism. There were 269 metabolites with differences in content between the eugenol intervention group and the thioacetamide group. Compared with the thioacetamide group, there were 101 metabolites with increased content and 168 metabolites with decreased content in the eugenol intervention group. These differential expressed metabolites suggest that eugenol intervention can correct inflammation damage, oxidative damage, and energy metabolism abnormalities caused by TAA. This study found through pathological section observation and metabolomics that eugenol has a protective effect on acute liver injury caused by thioacetamide, and the protective mechanism may be related to the antioxidant and anti-inflammatory effects of eugenol.