Integrated network pharmacology, molecular docking, and experiments in vivo and in vitro to explore the efficacy and potential mechanism of Chrysanthemum indicum L. extract against glioma

Chrysanthemum indicum L., a herbaceous perennial plant in the Asteraceae family, has been found to possess anti-inflammatory and anticancer effects. Jiawei Juming Decoction (JWJM), composed of Chrysanthemum indicum L., Senna tora (L.) Roxb., Forsythia suspensa (Thunb.) Vahl, Astragalus membranaceus (Fisch.) Bunge, etc., has been utilized in the clinical for treating glioma in China. However, as an important medicinal material, the efficacy of Chrysanthemum indicum L. against glioma have not been reported. In this study, a multi-faceted research approach encompassing in vitro and in vivo experimentation, network pharmacology, and molecular docking was employed to elucidate the anti-glioma effects of Chrysanthemum indicum L. extract (CIE) and the changes in related target proteins within Pathways in cancer. Active ingredients in CIE were identified using the TCMSP and ETCM databases. The GeneCards and DisGeNET databases were utilized to identify targets associated with glioma, followed by creating a Venn map to pinpoint common drug-disease targets. The STRING database was employed for analyzing protein-protein interaction (PPI) and screening key targets. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were performed for the enrichment analyses. Cytoscape software was utilized to create a compound-target-pathway network aimed at discovering important active components and potential mechanisms. The binding strength between the key target and CIE’s active compounds was verified via molecular docking simulation. The predicted results were confirmed by cell experiments, and the influence of CIE on glioma cell proliferation, migration, apoptosis, as well as antitumor activity were further evaluated through in vivo and in vitro experiments. The CIE contains 23 active compounds and 130 major targets, identifying nine key targets: ESR1, SIRT1, HSP90AA1, PTGS2, RELA, AR, NOS3, DNMT1, and GSK3B. The effectiveness of CIE treatment is primarily focused on the Pathways in cancer, as highlighted by GO and KEGG enrichment analysis. It was confirmed through protein expression that Androgen receptor (AR) is essential for glioma progression. Molecular docking showed that kaempferol, naringenin and luteolin strongly bound to AR. The influence of CIE on the target protein was confirmed through in vitro experiments. It has been demonstrated that CIE has the capacity to inhibit the proliferation and migration of C6 cells, whilst concomitantly inducing an increased rate of cell apoptosis. Experiments with animals revealed that CIE could initiate apoptosis in glioma cells and possessed anti-tumor properties. By influencing the relevant target protein (AR), the proliferation of C6 cells could be reduced by CIE, which also boosts their apoptosis. This study offers a robust evidential basis for the clinical application and commercial progression of CIE.