Multi-target modulation of the neuroinflammatory network by asiatic acid in ischemic stroke: Integrated network pharmacology, molecular docking, and in vitro validation 

Abstract

Asiatic acid (AA), a pentacyclic triterpenoid derived from Centella asiatica, has shown neuroprotective and anti-inflammatory activities, but the molecular network underlying its potential effects in ischemic stroke remains incompletely understood. Here, network pharmacology, protein–protein interaction (PPI) analysis, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, molecular docking, and in vitro experiments were integrated to define a multi-target neuroinflammatory framework for AA. A total of 194 AA-associated targets and 2,721 ischemic stroke-associated targets were identified, yielding 96 overlapping targets. PPI analysis prioritized IL-6, TNF, AKT1, and IL-1β as prominent hub candidates, while enrichment analysis linked the shared target network to hypoxia responses, inflammatory regulation, apoptosis, HIF-1 signaling, and TNF-related pathways. Molecular docking predicted structurally plausible interactions between AA and AKT1, TNF, IL-1β, and IL-6. In HT22 cells subjected to oxygen-glucose deprivation/reoxygenation (OGD/R), AA improved cell viability, reduced TNF-α, IL-6, and IL-1β release, and attenuated OGD/R-induced increases in Akt1, Tnf, Il1b, and Il6 mRNA. Collectively, these findings support a hypothesis in which AA modulates an interconnected neuroinflammatory and stress-response network rather than a single molecular target. IL-6, TNF, IL-1β, and AKT1 therefore emerge as experimentally supported candidate nodes for further target-validation studies and, with appropriate in vivo and clinical validation, as potential biomarker candidates. This integrated framework expands the therapeutic rationale for AA in ischemic stroke while defining specific targets and pathways for future mechanistic investigation.