Unraveling the therapeutic mechanisms of Huanglian Jiedu Tang in bacterial meningitis: A comprehensive approach integrating network pharmacology, molecular docking, and experimental validation.

This study aims to comprehensively elucidate the therapeutic mechanisms of Huanglian Jiedu Tang in bacterial meningitis via an integrated strategy combining network pharmacology, molecular docking, and experimental validation. Network pharmacology was utilized to screen for bioactive constituents, pivotal targets, and associated signaling pathways of HLJDT. The potent candidate Obacunone and its target MAPK14 were prioritized for verification. An in vitro bacterial meningitis cell model was established, followed by therapeutic interventions using Obacunone, the MAPK14 inhibitor VX-702, or their combination. Therapeutic effects were assessed using the CCK-8 assay, sodium ion permeability assays, TUNEL staining, RT-PCR, Western blot, and ELISA. Bioinformatics analysis of the GEO dataset GSE40586 identified 495 BM-related genes, 30 of which intersected with HLJDT targets. GO and KEGG enrichment analyses highlighted biological processes including antibiotic and glucocorticoid responses, as well as transcriptional misregulation and HIF-1 signaling pathways. Molecular docking simulations revealed that Obacunone possessed superior binding affinity to MAPK14. In the BM cell model, treatment with Obacunone or VX-702 alone significantly improved cell viability, attenuated the increased sodium ion permeability, inhibited apoptosis, and suppressed inflammatory cytokine levels. Notably, combination therapy exhibited the most potent synergistic effects. In a bacterial meningitis co-culture model, combined treatment with Obacunone and VX-702, as well as MAPK14 silencing, significantly restored the viability and migratory capacity of HMC3 cells while effectively attenuating early apoptosis. This study delineated the pharmacological underpinnings of HLJDT in BM treatment, identifying its active components and targets. Experimental validation confirmed the synergistic action of Obacunone and the MAPK14 inhibitor, providing a robust scientific foundation for future drug development and clinical strategies.