Objective To investigate the protective effect of puerarin (Pue) on acute liver failure (ALF)induced by d-galactosamine in mice and its mechanism. Methods Fifty Kunming mice were randomly divided into fivegroups. Two weeks before modeling, the Pue group, LY294002 group (LY group), and Pue + LY group were injected with300 mg/ kg Pue, 10 mg/ kg LY, or 300 mg/ kg Pue + 10 mg/ kg LY, respectively, into the caudal vein once daily. Thenormal control and model group rats were administered the same amount of sterile physiological saline. After the finaladministration and fasting for 24 h, the ALF model was established by intraperitoneal injection of galactose in all groupsexcept the control group which received the same amount of sterile normal saline. Serum levels of alanine aminotransferase(ALT), aspartate transferase (AST), and total bilirubin (TBil) were detected by a biochemical analyzer, and levels ofmalondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in liver tissues weredetected by kits. Hematoxylin and eosin staining, TUNEL staining, and western blotting were used to detect pathologicalchanges in the mouse liver tissue, apoptosis in liver cells, and the expression levels of P-Akt, P-glycogen synthase kinase(GSK)-3β, and cleaved caspase-3 protein, respectively. Results Compared with the control group, significant increaseswere detected in the number of apoptotic hepatocytes, serum ALT, AST, and TBil levels, the liver tissue MDA content,and cleaved caspase-3 protein expression levels in the model group ( P < 0. 05), while protein expression levels of SOD,GSH-Px, P-Akt, and P-GSK-3β were significantly decreased ( P < 0. 05). After treatment, all indices in the Pue groupwere significantly improved compared with the model group ( P < 0. 05), while there were no significant differencesbetween indices among the LY group, Pue+LY group, and model group ( P > 0. 05). Conclusions Puerarin may play aprotective role in the liver of mice with d-galactosaminoglycan-induced acute liver failure by activating the PI3K/ Akt signaling pathway, thereby reducing the degree of liver function damage.