Exploring neuroprotective effects of PP2 in ischemic stroke via bioinformatics and experimental validation

Ischemic stroke is a leading cause of mortality and disability worldwide, yet effective therapeutic options remain limited. In this study, bioinformatics analyses were used to identify potential therapeutic targets and small-molecule compounds for ischemic stroke. A mouse model of cerebral ischemia was subsequently used to validate their neuroprotective efficacy. Bioinformatics methods were used to analyze and identify key signaling pathways and hub genes associated with ischemic stroke. Additionally, the Connectivity Map (CMap) database was queried to identify potential small-molecule compounds for ischemic stroke treatment. Finally, a middle cerebral artery occlusion/reperfusion (MCAO/R) mouse model was employed to further evaluate the neuroprotective effects of the identified compounds. GO and KEGG pathway enrichment analyses revealed that key signaling pathways such as TNF, NF-κB, and IL-17 play crucial roles in ischemic stroke. PPI network analysis identified five hub genes-IL-1β, IL-6, ICAM-1, Jun, and Fos-all closely associated with neuroinflammatory responses. The small-molecule compound PP2, a selective Src kinase inhibitor, was identified by CMap database. In the MCAO/R mouse model, PP2 exhibited significant neuroprotective effects. It reduced infarct volume and brain edema and improved neurological function. Mechanistically, PP2 inhibited Src phosphorylation, thereby suppressing the NF-κB signaling pathway and reducing levels of pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6. This study identifies Src kinase as a promising therapeutic target for ischemic stroke and highlights the value of bioinformatics in drug discovery and mechanistic research.