Network Pharmacology Combined with Experimental Validation to Investigate the Mechanism of Huxinkang Tablets in Treating Atherosclerosis by Modulating Macrophages

Huxinkang tablets (HXKT) is the formula prescription of Traditional Chinese Medicine for treating atherosclerosis (AS), but its underlying mechanisms remain unclear. Network pharmacology and experimental verification were integrated to explore the therapeutic effects and key targets of HXKT in macrophage-associated genes in vulnerable AS plaques. Our findings indicated that a total of 75 candidate targets of HXKT against vulnerable AS were acquired. Furthermore, five core targets were identified including STAT3, HIF1A, EGFR, ESR1, and BCL2. KEGG pathway analysis demonstrated that these targets participated in the advanced glycation end product – receptor for advanced glycation end-product (AGE-RAGE) signaling pathway associated with diabetic complications, lipid metabolism, AS, and human cytomegalovirus infection. Molecular docking results indicated that the ESR1-miltirone complex exhibited the highest binding affinity. In vitro experiments demonstrated that HXKT inhibited oxidized low-density lipoprotein-induced macrophage injury, reversed abnormal expression of key targets, and reduced inflammation and lipid accumulation. Overexpression of STAT3 partially attenuated the protective effects of HXKT, confirming its role as a critical target while underscoring the multi-target mechanism of HXKT. In conclusion, HXKT stabilizes vulnerable atherosclerotic plaques by regulating macrophage function through a multi-component, multi-target mechanism, with STAT3 signaling playing a pivotal role.