关键词: 丹灯通脑软胶囊, 指纹图谱, 网络药理学, 药效物质基础, 作用机制, 缺血性脑卒中

Abstract: BACKGROUND: The Chinese medicine Compound Dandeng Tongnao Soft Capsule is characterized by the synergistic effect of multiple components, but traditional research methods are difficult to systematically reveal its complex mechanism of action. This experiment innovatively integrates high-performance liquid chromatography fingerprinting and network pharmacology methods. This integrated strategy can not only achieve a comprehensive evaluation of the material basis of traditional Chinese medicine, but also further elucidate the targets of traditional Chinese medicine, clarify its pathways and mechanisms, and provide a new research paradigm for elucidating the “multi-component-multi-target-multi-pathway” mechanism of traditional Chinese medicine prescriptions.
OBJECTIVE: To investigate the material basis and mechanism of action of Dandeng Tongnao Soft Capsules against ischemic stroke based on HPLC fingerprinting and network pharmacology. 
METHODS: The fingerprints of 10 batches of Dandeng Tongnao Soft Capsules were determined by high-performance liquid chromatography, and the common peaks in the fingerprints of Dandeng Tongnao Soft Capsules were identified by LCMS-IT-TOF technique. Common peak component targets of Dandeng Tongnao Soft Capsules were predicted by SwissTargetPrediction database (developed by Swiss Institute of Bioinformatics; function: prediction of potential targets based on the structure of compounds). Ischemic stroke-related targets were collected through the OMIM database (maintained by Johns Hopkins University, USA; function: to include human genetic diseases and disease-causing genes) and the GeneCards database (developed by the Weizmann Institute of Science, Israel; function: to integrate information on gene functions, disease associations and pathways). Drug-disease intersection targets were screened using the Venny 2.1.0 online tool to draw Venn diagrams. Cytoscape 3.10.1 software was used to construct a “drug-compound-target” interaction network to screen the potential components of Dandeng Tongnao Soft Capsules against ischemic stroke. Protein-protein interaction network analysis was performed through the STRING database (jointly developed by the European Molecular Biology Laboratory and other institutions, function: analysis of protein interaction networks and core targets) to obtain core targets. DAVID database (developed by National Institute of Allergy and Infectious Diseases, Function: Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis) was used to analyze the biological functions and pathways of the core targets of Dandeng Tongnao Soft Capsules against ischemic stroke, and ultimately to build a multi-dimensional network of “drug-component-target-pathway-disease.”
RESULTS AND CONCLUSION: (1) The fingerprint pattern of Dandeng Tongnao Soft Capsules was established, 24 common peaks were calibrated, the peaks were well separated, and the similarity between samples was greater than 0.9. Twenty-four peaks in Dandeng Tongnao Soft Capsule were identified. A total of 508 intersecting targets of components and diseases in Dandeng Tongnao Soft Capsule were obtained. (2) Protein-protein interaction analysis, GO function enrichment and KEGG pathway enrichment analysis revealed that Dandeng Tongnao Soft Capsules may exert anti-ischemic stroke effects through its active ingredients, geranodendronin, danshenone I, tanshenone IIA, ethyl ferulate, and tanshenaldehyde, which act on glyceraldehyde-3-phosphate dehydrogenase, serine/threonine protein kinase 1, and tumor necrosis factor, non-receptor tyrosine kinase, to regulate hypoxia-inducible factor-1 signaling pathway, advanced glycosylation end product-receptor signaling pathway, and phosphatidylinositol 3-kinase protein kinase B signaling pathway. To conclude, the organic integration of chemical analysis and bioinformatics prediction systematically explains the multidimensional mechanism of action of Dandeng Tongnao Soft Capsules. It not only provides a scientific basis for its clinical efficacy, but also, more importantly, establishes an integrated research framework of “fingerprinting-network pharmacology,” which provides a general methodological reference for the comprehensive evaluation of the material basis of traditional Chinese medicines and the analysis of their mechanism of action, and has important theoretical and practical value for the modernization of traditional Chinese medicines.

Key words: Dandeng Tongnao Soft Capsule, fingerprinting, network pharmacology, substance basis of efficacy, mechanism of action, ischemic stroke