<p>Sepia ink, a defensive secretion produced by cephalopods, is a traditional Chinese marine medicine. Although previous studies have demonstrated its efficacy in lead (Pb) removal, its hepatoprotective effects and the associated mechanisms remain poorly understood, limiting its broader therapeutic potential. This study evaluated its protective effects against Pb toxicity in mice. Sixty Kunming (KM) mice were divided into control, model, positive drug (dimercaptosuccinic acid, DMSA), and three experimental groups receiving 250, 500, and 1000 mg/kg sepia ink, respectively. Except for controls, all mice received drinking water containing 0.2% lead acetate (CH₃COO)₂Pb for 28 d. Concurrently, the three sepia ink groups and the DMSA group received corresponding oral doses. Pb levels in blood, liver, and bone tissues were measured by ICP-MS. Serum and liver samples were collected to assess liver function, key indicators of oxidative stress and antioxidant capacity, and to analyze endogenous metabolites using UPLC-QE-Orbitrap-MS. After a 4-week intervention, sepia ink significantly reduced Pb concentrations in the blood, liver and bone of treated mice. Moreover, it markedly reduced serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities, suggesting the hepatoprotective effects of sepia ink. Additionally, sepia ink alleviated oxidative stress by modulating key antioxidant indicators including superoxide dismutase (SOD) and glutathione peroxidase (GPX), and restored hepatic metabolic homeostasis through the regulation of 38 Pb-disrupted metabolites, primarily involved in steroid hormone biosynthesis, arachidonic acid metabolism, and ABC transporters pathways. These results demonstrate sepia ink’s dual therapeutic effects against Pb-toxicity: enhancing Pb removal while alleviating hepatic oxidative stress and restoring metabolic homeostasis.</p>