Blood-activating, depression-relieving formula alleviates post-stroke depression: mechanistic insights from network...

Post-stroke depression (PSD) is common and disabling, yet mechanism-based, multi-target therapies that jointly curb neuroinflammation and support cell survival are scarce. We evaluated a Blood-Activating, Depression-Relieving (BADR) herbal formula for effects on PSD-relevant molecular hubs and microglial phenotypes.

BADR constituents from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform were standardised and mapped to human protein targets. Target-disease interaction networks were assembled in Search Tool for the Retrieval of Interacting Genes/Proteins, clustered with Molecular Complex Detection, and functionally annotated via Kyoto Encyclopedia of Genes and Genomes Orthology-Based Annotation System (KEGG/GO). For experimental validation, BV2 microglia were activated with lipopolysaccharide (LPS; 24 h) and co-treated with BADR within a pre-established non-cytotoxic range; dexamethasone (1 μM) served as comparator. Outcomes included cytokines (IL-1β, TNF-α, IL-6; enzyme-linked immunosorbent assay), expression of selected nodes (EGFR, STAT3, JUN, PIK3CA, BCL2; quantitative real-time polymerase chain reaction/western blot), viability (Cell Counting Kit-8), and apoptosis (flow cytometry).

Network analysis highlighted two dense modules enriched for PI3K-AKT, JAK-STAT, and neuroactive-ligand signaling. Hubs included EGFR, AKT1, STAT3, JUN, PIK3CA, and BCL2, with EGFR, STAT3, PIK3CA, JUN, and BCL2 prioritised for cellular validation based on topology, pathway relevance, and compound-target connectivity. In BV2 cells, BADR attenuated LPS-induced IL-1β, TNF-α, and IL-6 surges, improved viability, and reduced total apoptosis, with directionally comparable effects to dexamethasone. Mechanistically, BADR down-regulated EGFR/JUN/STAT3/PIK3CA and restored BCL2 at transcript and protein levels.

By converging network-level predictions with microglial phenotyping, the formula exerts coordinated anti-inflammatory and pro-survival effects centred on the EGFR-STAT3-PI3K nodes in a PSD-relevant context. These data provide a mechanistic rationale for further phosphorylation-level and in vivo validation toward multi-target PSD therapeutics.