Objective To investigate the protective effect of Matrine injection on acute alcoholic liver injury in mice.Methods A total of 30 mice were randomly divided into blank control group, model group, positive control group (125 mg/kg of Bifendate Pills), high-dose Matrine injection group (2.8 mg/kg), and low-dose Matrine injection group (1.4 mg/kg), each group with 6 mice. Except for the blank control group and the model group, each group received tail vein injections at a dose of 10 ml/kg per day for 14 consecutive days. On day 15, the acute alcoholic liver injury model was established by gavage with 50% ethanol at a dose of 12 ml/kg in all the groups except for the blank control group. All the mice were fasted but not deprived of water. After 16 hours of the modeling, the blood was collected and the mice were sacrificed. HE staining was used to observe the pathological changes of the liver, and oil red O staining was applied to detect the hepatic fat deposition. In addition, serum levels of alanine transaminase (ALT), aspartate aminotransferase (AST), and total bilirubin (T-BIL), and hepatic levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), catalase (CAT) and triglyceride (TG) were detected.Results The hepatic steatosis index was lower in all other groups relative to that in the model group (P < 0.05). Compared with the model group, the serum levels of AST, ALT and T-BIL were reduced (P < 0.05), the hepatic levels of SOD, GSH and CAT were increased (P < 0.05), and the hepatic levels of MDA and TG were decreased in all other groups (P < 0.05). In addition, the number of hepatic lipid droplets was reduced in all other groups compared with the model group (P < 0.05).Conclusions Matrine injection can regulate the level of serum transaminases, improve the liver function, enhance the antioxidant capacity, and ameliorate the oxidative stress in mice, therefore exerting a protective effect on acute alcoholic liver injury.