Therapeutic effects of ginseng and its major components on ischaemic stroke in vitro and in silico.

BACKGROUND: Ischemic stroke (IS) poses a significant threat to human health, prompting the need for more effective treatments. Tianjiangxueshuantong (TJXST) pills, commonly used for cerebrovascular diseases, contain ginseng, whose therapeutic potential for IS remains underexplored. This study investigates the effects and mechanisms of ginseng in IS. METHODS: Initially, we established oxygen-glucose deprivation/reperfusion (OGD/R) models using bEnd.3 cells and glutamate-induced injuries in PC12 cells to assess the effects of ginseng extracts on cell activity using CCK8 assays. Subsequently, we employed network pharmacology in conjunction with the GEO database to identify the core active components of ginseng, key intervention targets in IS, and potential signaling pathways. We validated the interactions between the central targets and core components using molecular docking, and further confirmed these interactions at the molecular level through surface plasmon resonance (SPR). In addition, quantitative real-time PCR (qRT-PCR) and Western Blot (WB) analyses were performed to confirm mRNA and protein expression changes, respectively, and SRC knockdown experiments were conducted to assess its role in mediating the neuroprotective effects of ginseng. RESULTS: Our investigation identified arachidonate, kaempferol, celabenzine, suchilactone, ginsenoside Re, and ginsenoside Rg1 as the core active components of ginseng. We identified PTPN11, JAK2, PIK3R1, EGFR, PIK3CA, PIK3CB, SRC, and PTK2 as hub targets of ginseng intervention in IS. Molecular docking analyses confirmed the binding of the core component with the hub target. SPR assays demonstrated that the major active components of ginseng exhibited measurable binding to the core target, SRC. qRT-PCR and WB assays revealed changes in the expression of core targets in both cell models. In addition, SRC knockout experiments abolished the protective effects of ginseng extract, demonstrating that SRC is a core target of ginseng extract. CCK8 and ELISA experiments validated the increase in cell viability and reduction in inflammatory factor including TNF-α (Tumor Necrosis Factor-alpha), IL-1β (Interleukin-1 beta), VEGF (Vascular Endothelial Growth Factor), Ang-1(Angiopoietin-1) release mediated by the identified active ingredients. DISCUSSION: This study elucidates the molecular mechanisms by which ginseng exerts its therapeutic effects on IS. The identification of key active ingredients and their targets offers new insights into the development of effective treatments for IS.