BACKGROUND: Senescence of vascular endothelial cells (ECs) plays a critical role in cardiovascular diseases. Galangin (3,5,7-Trihydroxyflavone) is a flavonoid derived from plants and medicinal herbs with various biological activities. However, its effect on endothelial cell senescence and the underlying molecular mechanisms are poorly characterised.
PURPOSE: This study aimed to evaluate the therapeutic effects of Galangin on endothelial cell senescence and vascular aging, providing a theoretical basis for its clinical application.
METHODS: Human aortic endothelial cells and mice were treated with d-galactose to induce senescence. The expression levels of aging markers β-galactosidase, P21, P53, and γ-H2AX were detected using western blot. Cell proliferation was determined using CCK-8 assays. Network pharmacology, molecular docking, and molecular dynamics simulations were used to identify Galangin downstream targets. MST, CETSA, and DARTS assays were used to determine the binding of Galangin to SMAD3 protein. ROS levels were measured using flow cytometry. JC-1 staining was used to determine mitochondrial membrane potential. Mitophagy-related markers Parkin, PINK1, LC3, and P62 were detected using western blot and immunohistochemistry. ChIP assay was used to determine p-SMAD3's effect on the downstream genes PINK1 and LC3.
RESULTS: Our pioneering study reveals a novel protective role of Galangin in vascular aging, primarily through a newly identified mechanism: activating mitophagy by targeting SMAD3. Notably, Galangin directly binds to SMAD3 and promotes the transcriptional regulation of PINK1 and LC3.
CONCLUSIONS: Our findings demonstrate the protective effect of Galangin in vascular aging and hold promising prospects for both clinical application and future research.
