Exploring the pro-angiogenic constituents of Lagopsis supina and their mechanisms via transcriptome analysis,...

Lagopsis supina is a traditional Chinese medicinal herb used for activating blood circulation and nourishing the blood. This study aims to investigate the pro-angiogenic constituents of L. supina and their mechanisms of their action. 30 compounds derived from the active fraction LS-D (L. supina 60% ethanol-water extract) were screened, of which 15 showed angiogenesis-promoting activity, were screened in a PTK787-induced vascular injury zebrafish model. The active constituents were primarily phenolic acids and phenylpropanoids, among which compounds LS21 (vanillic acid) and LS22 (syringic acid) exhibited the most potent pro-angiogenic effects. These constituents (LS21 and LS22) were selected for further investigation of mechanisms of promoting angiogenesis. Transcriptomic analysis followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment revealed differentially expressed genes and predicted involved pathways. Molecular docking simulated interactions between compounds and key pathway proteins, and RT-qPCR validated gene expression patterns. Mechanistic studies integrating transcriptomics, molecular docking, and RT-qPCR revealed that both LS21 and LS22 downregulated immune-related cell adhesion molecules, while upregulating cdh5 and pecam1a. Additionally, LS21 specifically downregulated the signaling molecules smad2 and smad3a, and upregulated the transcriptional corepressors skila and ncor1. In contrast, LS22 downregulated extracellular matrix-related adhesion molecules and itga2b, while upregulating thbs1a and integrin itgb3b. In conclusion, the L. supina constituents LS21 and LS22 that are primarily phenolic acids and phenylpropanoids, exert their angiogenesis-promoting effects mainly via modulating cell adhesion and suppressing TGF-β signaling pathway.