AIM:  To investigate the potential mechanism of licorice on liver injury induced by Semen Strychni based on network pharmacology, molecular docking combined with animal experiments, providing an effective strategy for prevention and treatment of liver injury induced by Semen Strychni. METHODS: Firstly, the active ingredients of Semen Strychni and licorice were obtained through the ETCM, TCMSP database and analysis platform, CTD database and literature supplementation. Then, the potential toxic ingredients of Semen Strychni were further screened based on the SwissADME platform, and the targets corresponding to the active ingredients were predicted through the SwissTargetPrediction platform. By using the GeneCards and OMIM databases to collect DILI-related targets, the potential targets for licorice to alleviate liver injury caused by Semen Strychni were obtained. By constructing the active ingredient-target network, the core ingredients of licorice in alleviating liver injury caused by Semen Strychni were screened. The key targets obtained were used to construct and analyze the protein-protein interaction networks (PPI) through the STRING database and Cytoscape 3.9.0 software. The potential targets were subjected to GO and KEGG pathway enrichment analysis with the aid of the DAVID database, and constructed a network of active ingredient-target-pathway. Molecular docking study was approved for the core targets and the active ingredients by using Schrodinger 2023-1 software, and the visualization operation was conducted through Pymol. Finally, the regulatory effect of licorice on the key pathway of liver injury caused by Semen Strychni was validated by establishing a rat model of liver injury induced by Semen Strychni. RESULTS: After screening, 6 potential toxic components of Semen Strychni and 104 corresponding targets, 89 active components of licorice and 347 corresponding targets, and 3 200 DILI targets were obtained. A total of 23 intersection targets were obtained through Venn analysis. By constructing the active ingredient-target network, it was found that the main core ingredients were 7-methoxy-2-methyl isoflavone, medicarpin, shinpterocarpin, quercetin, formononetin and isoliquiritigenin. The PPI network indicated that the core targets were protein kinase B1 (AKT1), epidermal growth factor receptor (EGFR), human epidermal growth factor receptor 2 (ERBB2), glycogen synthase kinase 3 beta (GSK3B), kinase insert domain receptor (KDR) and Janus kinase 2 (JAK2). A total of 39 relevant pathways were enriched in KEGG (P<0.01), among which the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (AKT) (PI3K-AKT) signaling pathway, which has been confirmed and ranks first in enrichment, and was closely related to liver injury. Molecular docking results showed that the core components have good binding ability with the core targets. In vivo animal experiments demonstrated that, compared to the model group, licorice significantly reduced the liver index (P<0.01), serum levels of alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), indirect bilirubin (IBIL), and total bilirubin (TBIL) in rats with liver injury, while increasing total protein (TP) levels (P<0.05 or P<0.01). Additionally, licorice alleviated congestion in the central veins and hepatic sinusoids, improved the alignment of hepatocytes, and reduced inflammatory cell infiltration. Furthermore, licorice significantly decreased the levels of malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in the liver tissue of injured rats, while elevating the levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) (P<0.01). It also markedly downregulated the phosphorylation levels of PI3K and AKT (P<0.01). CONCLUSION: Licorice has multi-component and multi-target properties in the treatment of liver injury induced by Semen Strychni, which may play a hepatoprotective role by inhibiting the activation of PI3K-AKT signaling pathway.