Guanxinshu attenuates atherosclerosis by suppressing Galectin-3-mediated NLRP3 inflammasome activation

Atherosclerosis (AS) is a chronic inflammatory vascular disease strongly associated with the activation of the NLRP3 inflammasome. Studies have shown that Galectin-3 (Gal-3) promotes NOD-like receptor family pyrin domain-containing 3 (NLRP3) activation. To investigate whether Guanxinshu (GXS) alleviates AS by modulating Gal-3/NLRP3 inflammasome-associated signaling. ApoE^-/- mice fed a high-fat diet for 12 weeks were used to establish an AS model. Mice were divided into five groups: control, model, low-dose GXS (12.5 g/kg/day), high-dose GXS (25 g/kg/day), and simvastatin. Treatments were administered via oral gavage for 5 consecutive weeks. Human coronary artery endothelial cells (HCAECs) were treated with oxidized LDL (ox-LDL) to establish an injury model, followed by intervention with GXS-containing serum or the Gal-3 inhibitor TD139. Oil Red O staining, HE staining, immunohistochemistry, Western blotting, ELISA, immunofluorescence, co-immunoprecipitation, and molecular docking were used for comprehensive assessment. GXS significantly reduced aortic plaque area and lipid levels; it decreased IL-6, IL-18, and TNF-α levels in both serum and aorta. GXS suppressed the expression of Gal-3, NLRP3, ASC, caspase-1, and IL-1β proteins, and reduced Gal-3/NLRP3 co-localization. In vitro, GXS-containing serum improved HCAECs viability, reduced inflammatory cytokines, disrupted the interaction between Gal-3 and NLRP3, and reduced ox-LDL-induced NLRP3 inflammasome-associated protein expression, with effects comparable to TD139. Molecular docking predicted potential binding between representative candidate components of GXS and Gal-3, as well as potential binding affinity between Gal-3 and NLRP3 proteins. This study demonstrates for the first time that GXS mitigates vascular inflammation and slows AS progression by modulating Gal-3/NLRP3 inflammasome-associated signaling and reducing NLRP3 inflammasome-associated protein expression.