Integrative multi-Omics and network pharmacology reveal angiogenesis promotion by Quan-Du-Zhong Capsule via VEGFA/PI3K-Akt pathway

Ethnopharmacological relevance: Quan-du-zhong capsule (QDZ), derived from the whole plant extract of Eucommia ulmoides Oliv., is a traditional Chinese herbal medicine used in treating vascular-related diseases, including hypertension and osteoporosis. Despite its established uses, its pro-angiogenic effects and underlying mechanisms require further investigation. Aim of this study: This study aims to investigate the pro-angiogenic effects of QDZ and explore the underlying mechanisms. Materials and methods: The chemical compositions of QDZ, including its absorbed prototypes in rats, were analyzed using UHPLC-Q Exactive-Orbitrap-MS. The pro-angiogenic activities of QDZ were evaluated in human umbilical vein endothelial cells (HUVECs) through various assays, including CCK-8, migration, scratch, tubule formation, and 3D sprouting assays. Additionally, the pro-angiogenic effects of QDZ were further assessed in vivo through the matrigel plug assay and a hindlimb ischemia-reperfusion model, with three-dimensional blood flow visualized via micro-CT. A comprehensive approach involving network pharmacology, molecular docking, transcriptomics, and proteomics was utilized to explore the pro-angiogenic mechanism of QDZ, with validation by Western blot analysis. Results: QDZ significantly promoted the proliferation, migration, and tubule formation of HUVECs. The matrigel plug assay further confirmed its pro-angiogenic potential. In vivo, QDZ-treated mice displayed enhanced vascular distribution and faster blood flow recovery post-ischemia-reperfusion. Chemical analysis identified 49 compounds in QDZ, with 16 absorbed prototypes detected in rat plasma. Mechanistic investigations through network pharmacology, transcriptomics, and proteomics suggested that QDZ's pro-angiogenic effects were mediated through the VEGFA/PI3K-Akt signaling pathway, with increased phosphorylation of angiogenesis-related proteins such as PI3K, Akt, FAK, and Src. Conclusions: This study demonstrates that QDZ promotes angiogenesis via activating the VEGFA and its downstream PI3K-Akt signaling pathway, shedding light on the mechanisms that underpin its traditional medicinal use in vascular health.