Discovering a Multi-Component Combination against Vascular Dementia from Danshen-Honghua Herbal Pair by Spectrum-Effect Relationship Analysis

The Danshen-Honghua (DH) herbal pair exhibits a synergistic effect in protecting the cerebrovascular system from ischemia/reperfusion injury, but the therapeutic effect on vascular dementia (VaD) has not been clarified, and the main active ingredient group has not been clarified. In this work, the chemical constituents in DH herbal pair extract were characterized by UHPLC-QTOF MS, and a total of 72 compounds were identified. Moreover, the DH herbal pair alleviated phenylhydrazine (PHZ)-induced thrombosis and improved bisphenol F (BPF)- and ponatinib-induced brain injury in zebrafish. Furthermore, the spectrum-effect relationship between the fingerprint of the DH herbal pair and the antithrombotic and neuroprotective efficacy was analyzed, and 11 chemical components were screened out as the multi-component combination (MCC) against VaD. Among them, the two compounds with the highest content were salvianolic acid B (17.31 +/- 0.20 mg/g) and hydroxysafflor yellow A (15.85 +/- 0.19 mg/g). Finally, we combined these 11 candidate compounds as the MCC and found that it could improve thrombosis and neuronal injury in three zebrafish models and rat bilateral common carotid artery occlusion (BCCAO) model, which had similar efficacy compared to the DH herbal pair. This study provides research ideas for the treatment of VaD and the clinical application of the DH herbal pair.