Yindan Xinnaotong Soft Capsule improves post-ischemic stroke recovery by regulating CREB/BDNF-mediated synaptic plasticity and CREB/VEGFA-mediated angiogenesis.

BACKGROUND: Ischemic stroke is a leading cause of disability and cognitive impairment. Yindan Xinnaotong Soft Capsule (YD), a traditional Chinese patent medicine, is clinically utilized for cardio-cerebrovascular diseases, yet its potential roles in post-stroke recovery remains unclear. PURPOSE: To elucidate the protective effects and underlying mechanisms of YD on ischemic post-stroke recovery. METHODS: Middle cerebral artery occlusion/reperfusion (MCAO/R) and oxygen-glucose deprivation/reperfusion (OGD/R) models were utilized to investigate the effects of YD on post-stroke recovery. Cerebral atrophy (TTC staining), histopathology (H&E, Nissl, and Golgi staining), synaptic ultrastructure (transmission electron microscopy, TEM), and behavioral tests were assessed. RNA-seq and network pharmacology analysis were utilized to reveal the underlying mechanisms, and verified in brain tissues, 2D monolayer cell, and 3D neurovascular unit (NVU) spheroids. Molecular docking was performed to screen the potential active ingredients and verified by immunofluorescence and cellular thermal shift assay. RESULTS: YD significantly alleviated post-stroke brain atrophy volume, neurological deficit score, cognitive/motor deficit, and pathological structure and hippocampal synaptic ultrastructure damage. A total of 535 differentially expressed genes analysis revealed that CREB signaling with the key targets of CREB, BDNF, and VEGFA was the central mechanism. The synaptic plasticity marker of synaptophysin and angiogenesis marker of CD31 were upregulated in brain tissues and 3D NVU spheroids mediated by CREB/BDNF and CREB/VEGFA pathway, respectively. Co-treatment with the CREB inhibitor 666-15 attenuated YD's effects on CREB phosphorylation. Salvianolic acid B (from Salvia miltiorrhiza Bunge) and Kaempferol-3-O-rutinoside (from Ginkgo biloba leaf) were identified and validated as primary active ingredients with high CREB affinity. CONCLUSION: Our study demonstrated that YD promoted post-ischemic stroke recovery by promoting CREB/BDNF-mediated synaptic plasticity and CREB/VEGFA-mediated angiogenesis, highlighting its therapeutic potential for treating long-term cerebral ischemia injury.