Integrating multi-omics and network pharmacology in a systems approach: a mechanism study of ginseng-derived panaxadiol saponins against immune-mediated aplastic anemia.

ETHNOPHARMACOLOGICAL RELEVANCE Panax ginseng C.A. Mey. is traditionally utilized to "tonify Qi and replenish Blood," particularly in managing anemia-like syndromes. Panaxadiol saponins (PND) represent a standardized bioactive fraction from ginseng that embodies these historical properties. While PND is currently in Phase II clinical trials for aplastic anemia (AA), its systemic pharmacological mechanisms remain to be fully elucidated.

Aim

OF THE STUDY This study aimed to evaluate the therapeutic efficacy of PND against immune-mediated AA, and delineate its potential systemic regulatory mechanisms involving the NLRP3-related signaling and gut-bone marrow crosstalk.

Methods

An immune-mediated AA mouse model was established. Network pharmacology, 4D-DIA quantitative proteomics and 16S rDNA sequencing were integrated to identify responsive molecular pathways and candidate targets. Predicted target interactions were characterized via molecular dynamics (MD) simulations and further validated by cellular thermal shift assay (CETSA). Functional validation was performed in an LPS-induced primary bone marrow nucleated cells (BMNC) injury model using the NLRP3-specific inhibitor MCC950, to investigate the functional involvement of the NLRP3 axis in PND-mediated cytoprotection.

Results

PND partially restored peripheral blood counts and ameliorated CD4+/CD8+ T-cell imbalances in AA mice. Integrative analysis identified the NOD-like receptor (NLR) signaling pathway as a candidate key responsive node, with MD simulations and CETSA characterizing potential biophysical interactions between ginsenosides and the chaperone HSP90AA1. Concurrently, PND treatment improved intestinal barrier integrity and enriched beneficial microbiota, changes that were associated with attenuated systemic endotoxemia. In vitro functional rescue assays further demonstrated that NLRP3 signaling axis is functionally involved in the anti-inflammatory and cytoprotective effects of PND on BMNCs.

Conclusion

PND facilitates hematopoietic recovery, an effect associated with the suppression of the NOD/NLRP3 inflammatory axis and a reduced systemic inflammatory burden. These findings suggest that PND holds potential as an adjunctive supportive strategy for immune-related cytopenias.