Abstract
Vascular aging constitutes a predominant risk factor for cardiovascular pathologies. Traditional Chinese Medicine (TCM) employs various formulations to mitigate age-related vascular dysfunction, among which Yiqihuoxue decoction (GSC) is clinically utilized for managing cardiovascular conditions in elderly patients. Our prior work demonstrated GSC’ s capacity to delay vascular aging in mice models and attenuate senescence in vascular endothelial cells, though its mechanistic basis remained unresolved. To address this, we conducted multi investigation combining Ultra-Performance Liquid Chromatography (UPLC), network pharmacology, and molecular dynamics (MD) simulations. UPLC identified 130 bioactive compounds in GSC, while integrative analyses (mass spectrometry, literature mining, and target prediction) revealed 792 putative targets. Intersection with 2,539 vascular aging-associated targets yielded 422 shared candidates, suggesting GSC’ s polypharmacological potential. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses highlighted autophagy-related pathways, notably PI3K/Akt and SIRT1 signaling. Cellular validation experiments in senescent human umbilical vein endothelial cells (HUVECs) demonstrated that GSC restored cell morphology and reduced SA-β-gal activity (p < 0.05). GSC alleviated G0/G1 phase cell cycle arrest, restored mitochondrial membrane potential, and suppressed ROS accumulation. Autophagy profiling indicated that GSC promoted autophagic flux, as evidenced by increased LC3B puncta formation in immunofluorescence assays and autophagosome accumulation observed via transmission electron microscopy. Mechanistically, GSC exerted anti-senescence effects via coordinated regulation of the SIRT1-autophagy axis and PI3K/AKT pathway inhibition. Western blotting confirmed dose-dependent upregulation of SIRT1 and downregulation of p-PI3K/p-AKT (p < 0.05), consistent with network pharmacology predictions. This study established GSC as a multi-component, multi-target intervention against vascular aging, with autophagy modulation serving as a central mechanism. These findings will provide a theoretical foundation for developing GSC-based therapies targeting age-related cardiovascular diseases. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1038/s41598-026-44263-4.