Objective:To explore the molecular mechanisms of protective effects of glutathione(GSH)in mice with acetamino-phen-induced acute liver injury.Methods:120 mice were randomly divided into 3 groups:Control,APAP,GSH,with 40 in each group.After establishing the model,mice in the GSH groups were intraperitoneally injected with GSH(0.40 g/kg)for 3 days.Subsequently,liver tissue samples were collected.SWATH combined with liquid chromatography-tandem mass spectrometry was used to analyze the protein expression profile,and the differentially expressed proteins between APAP group and Control group,GSH group and APAP group were screened out.GO functional annotation and KEGG pathway enrichment analysis were performed for the above differential proteins.Protein interaction network(PPI)was constructed by String database and Cytoscape software,and the key proteins were verified by im-munohistochemical staining.Results:A total of 310 differentially expressed proteins were screened out between APAP group and Control group.A total of 172 differentially expressed proteins were screened out between the GSH group and the APAP group.The enrichment a-nalysis of the above differentially expressed proteins showed that the differential proteins were mainly enriched in the functions and pathways of nitrogen metabolism,fatty acid metabolism,and organic acid metabolism.Further analysis identified 17 differentially ex-pressed proteins between APAP group vs.Control group and GSH group vs.APAP group.Anxa5,Rgn,Tagln,Vim and Usp5 may be re-lated to apoptosis.Anxa5 and Rgn protein expression was higher in the APAP group than in the Control group(P＜0.05).In contrast,Anxa5 protein expression decreased and Rgn protein expression increased in the GSH group compared to the APAP group(P＜0.05),there was no statistically significant difference in Tagln,Vim,and Usp5(P＞0.05).Conclusion:GSH protects mice from APAP-in-duced liver injury by inhibiting cell apoptosis.Anxa5,Rgn,Tagln,Vim,and Usp5 are potential targets.