Unveiling the mechanisms of yanhusuo's therapeutic effects in neuropathic pain through network pharmacology, single-cell RNA sequencing, and molecular docking.

Background: Current therapeutic strategies for neuropathic pain (NP) encompass pharmacological agents, physical modalities, psychological support, and interventional procedures, which aim to mitigate inflammation, enhance vascular perfusion in afflicted regions, and modulate immune responses. However, the heterogeneity of NP pathogenesis and individual variability often lead to inconsistent treatment outcomes.

Methods: An integrative network pharmacology framework was employed to elucidate the mechanistic basis of Yanhusuo in NP management. NP patients were categorized via unsupervised clustering, followed by single-cell sequencing and cell-cell communication analysis to identify immune cell interactions. Active compounds and targets of Yanhusuo were identified using the Traditional Chinese Medicine Systems Pharmacology (TCMSP) and SwissTargetPrediction databases. Network pharmacology tools, including Cytoscape, facilitated the construction of protein-protein interaction (PPI), compound-target-disease, and compound-target-pathway networks. Topological analyses identified core targets and pathways, while the Database for Annotation, Visualization and Integrated Discovery (DAVID) bioinformatics platform was used for functional enrichment analysis. Finally, molecular docking analysis was conducted to evaluate ligand-receptor binding affinities.

Results: Nine bioactive compounds and 53 NP-associated targets were identified in Yanhusuo. PPI analysis suggests that ACTB, PPP1CA, ERK1, and PTEN may be the hub nodes with maximal centrality. KEGG pathway enrichment highlighted the focal adhesion pathway as pivotal in Yanhusuo's anti-NP activity. Molecular docking suggests that there may be strong binding interactions between key compounds and hub targets (e.g. binding energy<-6.5 kcal/mol).

Conclusions: This work systematically maps Yanhusuo's multi-target, multi-pathway therapeutic landscape in NP, offering a strategic foundation for mechanistic research and drug discovery. The identified bioactive candidates represent promising candidates for NP therapeutics.