Multi-pathway mechanisms of Liujunzi decoction in promoting glioma apoptosis and reversing drug resistance: network pharmacology and experimental validation.

BACKGROUND: Glioma is a highly aggressive brain tumor with a poor prognosis. Recent studies have demonstrated the anti-tumor potential of Liujunzi decoction (LJZD), but its specific effects and mechanisms in glioma remain unclear. This study aimed to elucidate the anti-glioma effects of LJZD and explore its underlying therapeutic mechanisms. METHODS: The active ingredients of LJZD were retrieved from the TCMSP database and screened using ADME analysis. Potential targets of both the ingredients and glioma were retrieved from various databases. The overlapping targets were identified as LJZD's therapeutic targets for glioma. To elucidate the biological functions and potential association among the overlapping targets, protein-protein interaction (PPI) network analysis, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted. Molecular docking was employed to evaluate the binding affinities between core targets and LJZD's main active ingredients. The inhibitory effect of LJZD on U251 glioma cells was validated by in vitro experiments. RESULTS: In total, we identified 76 active ingredients and 752 potential targets of LJZD, while 1,456 glioma-related targets were collected from databases, yielding 174 overlapping targets through intersection. PPI network analysis revealed 40 hub targets, with TP53, AKT1, HIF1A, TNF and IL6 ranking as the top five core targets based on degree. GO and KEGG enrichment analyses demonstrated that LJZD's anti-glioma effects are mediated through genes related to apoptosis and key pathways including platinum drug resistance, EGFR tyrosine kinase inhibitor resistance, and PD-LI expression and PD-1 checkpoint pathway in cancer. Molecular docking confirmed good to strong binding affinities between LJZD's main active ingredients (glabridin, pinocembrin and chrysophanol) and all top 10 core targets. In vitro experiments showed that glabridin inhibited U251 cell viability and promoted apoptosis. Glabridin significantly downregulated phosphorylation levels of AKT, STAT3 and c-Jun, while reducing expression of c-Myc and HIF-1α in U251 cells. CONCLUSION: LJZD may exert anti-glioma effects by inducing tumor cell apoptosis and overcoming drug resistance, potentially serving as an adjuvant to enhance conventional therapies. Additionally, its active ingredients (e.g., glabridin, pinocembrin and chrysophanol) provide novel leads for developing small-molecule therapeutics.