Uncovering the mechanism of the Shenzhi Jiannao formula against vascular dementia using a combined network pharmacology approach and molecular biology

Shenzhi Jiannao formula (SZJNF) is a herbal prescription which is used for detoxification, dredging collaterals, and activating blood circulation and Qi flow in traditional Chinese medicine. SZJNF is a clinical effective prescription for the treatment of vascular dementia (VD) first formulated based on the classical theory of traditional Chinese medicine, but its anti-VD mechanism remains ambiguous. The aim of this study was to elucidate the multi-target mechanisms of SZJNF against VD using a network pharmacology approach and verify its effects through biological experiments. We utilized network pharmacology-based prediction and molecular docking techniques to uncover the potential micro-mechanism of SZJNF against VD. We identified active components and potential targets, and performed network analysis, functional annotation, and pathway enrichment analysis. Subsequently, glutamate-induced PC12 cells and VD rats were used to verify the molecular mechanisms of SZJNF. Seventeen active compounds were identified in SZJNF rat plasma; moreover, 773 predicted targets and 1544 VD-related targets were found. Various networks, including the PPI, herb-compound-target, and compound-target-pathway network were constructed. A total of 188 shared targets were identified by network topological analysis, which were closely associated to the anti-VD effects of SZJNF. They were also enriched in various biological processes through hypoxia reaction, promotion of cell proliferation, inhibition of apoptosis, neuroactive ligand-receptor interaction, and calcium signaling pathway, as evaluated by the analysis of advanced functions and pathways. SZJNF components docked well with the key targets. Treatment with SZJNF promoted cell proliferation, ameliorated apoptosis and oxidative stress injury, and improved neurological and cognitive abilities. This study comprehensively demonstrated the multi-target mechanisms of SZJNF in VD using network pharmacology and molecular biology experiments. This provides evidence for further mechanistic studies and for the development of SZJNF as a potential treatment for patients with VD.