Pushen capsule treatment promotes functional recovery after ischemic stroke

Background: As a leading cause of long-term disability, ischemic stroke urgently needs further research and drug development. Pushen capsule (Pushen) has been commonly applied in clinical treatment for relieving headaches, dizziness, and numbness. However, the effects of Pushen on ischemic stroke have not been revealed yet. Purpose: To assess the efficiency of Pushen in ischemic stroke and identify its potential therapeutic targets and active ingredients for treating ischemic stroke. Study design and methods: Behavioural experiments, Triphenyltetrazolium chloride (TTC) staining, Magnetic resonance imaging (MRI), and immunofluorescence staining were performed to examine the efficiency of Pushen in stroke model mice. The potential mechanism and active ingredients of Pushen were assessed by transcriptome, 16S rDNA sequencing, metabonomics, and network pharmacology. Finally, the targets were validated by RT-PCR, chromatin immunoprecipitation (ChIP), ELISA, and molecular docking methods. Results: Pushen had several effects on stroke model mice, including reducing the infarct volume, improving the blood-brain barrier (BBB), and promoting functional restoration. Furthermore, the network pharmacology, LC-MS/MS, and molecular docking results revealed that tricin, quercetin, luteolin, kaempferol, and physcion were identified as the key active ingredients in Pushen that treated ischemic stroke. Mechanistically, these key in-gredients could bind with the transcription factor c-Myc and thereby regulate the expression of Adora2a, Drd2, and Ppp1r1b, which are enriched in the cAMP signaling pathway. Additionally, Pushen improved the gut microbiota dysbiosis and reduced inosine levels in feces and serum, thereby reducing Adora2a expression in the brain. Conclusions: Our study confirmed that Pushen was effective for treating ischemic stroke and has promising clinical applications.