Abstract: To observe the protective effect of peony seed oil on alcohol induced acute liver injury in mice, 60 Kunming mice were randomly divided into 6 groups: normal group (CK), model group (M), positive group (Y), peony seed oil low (D), medium (Z) and high (G) dose group. After 14 days of continuous administration, except the normal group, all mice were given 50% ethanol 19 mL/kg·BW at one time to induce acute liver injury and changes in mouse body weight and blood glucose were monitored. After 6 hours of alcohol gavage, mouse serum and liver tissue were collected to measure blood clearance indicators and prepare liver tissue slices. The results showed that during the gavage period, the body weight of all groups of mice showed an upward trend, and there was no significant difference (P>0.05), indicating that peony seed oil had no significant effect on the basal body weight of mice. After 6 hours of alcohol gavage, the blood glucose level of mice in the G group was the lowest (6.8 mmol/L), significantly lower than that of the M group (P<0.05), and not significantly different from the Z group (P>0.05). All dose groups of peony seed oil could reduce the AST, ALT enzyme activity, blood glucose, liver coefficient, TC, TG, MDA content, and increase the T-CO₂, GSH content, and T-SOD enzyme activity in experimental mice. The AST, ALT, TC, TG, and T-CO₂ levels in Z group were 196 U/L, 77 U/L, 2.67 mmol/L, 1.21 mmol/L and 23.1 mmol/L, respectively, which were significantly different from M group (P<0.01). The T-CO₂ level in Z group was significantly higher than that in G group (P<0.05), while other indicators showed no significant differences compared to G group (P>0.05). Liver tissue slices showed that medium doses of peony seed oil could significantly alleviate liver tissue lesions. All the results illustrated that peony seed oil had certain protective effect on alcohol induced acute liver injury in mice, with the medium dose group showing better efficacy.