Investigating the neuroprotective effects and the potential mechanisms of β-amyrenone in cerebral ischemic/reperfusion injury based on pharmacology network, bioinformatics, and molecular docking simulation

Cerebral ischemic/reperfusion injury (CIRI) can cause secondary injury to the neurons, which remains unresolved in the clinic. β-amyrenone harbors anti-inflammatory effects. The present study aims to explore the neuroprotective effects and the underlying mechanisms of β-amyrenone in CIRI based on network pharmacology, bioinformatics, and molecular docking simulation. Network pharmacology was used to screen the potential targets of the main bioactive ingredients of Euonymus fortunei in acute ischemic stroke (IS), and the disease-targets-drugs model was constructed using the Cytoscape software. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed by the Database for Annotation, Visualization, and Integrated Discovery (DAVID). The scores of the top 10 hub targets and the interaction degrees of the main bioactive ingredients were quantified by four algorithms (Degree, Betweenness Centrality, Closeness Centrality, and Neighborhood Connectivity) based on the topological structures of the constructed networks. The potential interactions between β-amyrenone and potential hub targets were predicted by molecular docking simulation. CIRI model was established through oxygen-glucose deprivation/reoxygenation (OGD/R) in neural cells. The neural protective effects of β-amyrenone were tested by cell counting kit-8 assay, enzyme-linked immunosorbent assay and flow cytometry. Reverse-transcription quantitative polymerase chain reaction and western blot were used to detect the influences of β-amyrenone on the levels of screened targets. Interleukin 1β (IL-1β), prostaglandin-endoperoxide synthase 2 (PTGS2), mitogen-activated protein kinase 3 (MAPK3), and estrogen receptor 1 (ESR1) might be the potential hub targets of the main bioactive targets of Euonymus fortunei in acute IS, and β-amyrenone was the core bioactive ingredient of Euonymus fortunei. Results of GO and KEGG pathway enrichment analysis displayed that β-amyrenone might modulate inflammation-related activities in acute IS. β-amyrenone could bind to the amino acid residues of IL-1β, PTGS2, MAPK3, and ESR1 with high affinity. Besides, β-amyrenone treatment increased neural cell viability, reduced the release of inflammatory cytokines, and protected from apoptosis under the OGD/R context. β-amyrenone treatment in neural cells with OGD/R exposure could influence the mRNA and protein levels of IL-1β, PTGS2, MAPK3, and ESR1. β-amyrenone exerts the neuroprotective effects against cerebral ischemic/reperfusion injury, which may be achieved by targeting IL-1β, PTGS2, MAPK3, or ESR1.