Huaban Tongmai decoction attenuates ferroptosis-associated vascular inflammation in carotid artery atherosclerosis with involvement of the p53-Parkin mitophagy pathway

Background Carotid artery atherosclerosis (CAS) is a chronic inflammatory vascular disease in which ferroptosis—an iron-dependent, oxidative form of regulated cell death—has been implicated in endothelial injury and vascular inflammatory responses. Despite growing evidence for the therapeutic potential of Huaban Tongmai decoction (HBTMD) against CAS, existing studies have not clearly elucidated its specific molecular mechanisms. This study focused on exploring and elucidating the mitigating mechanism of HBTMD against CAS. Methods An in vivo ApoE-/- mouse model of CAS was constructed via high-fat diet (HFD) combined with carotid ligation, and an ox-LDL-induced endothelial injury model using human umbilical vein endothelial cells (HUVECs). The chemical components of HBTMD were screened by UHPLC-MS/MS, and its marker constituents were quantified. Integrative proteomics and untargeted metabolomics identified key DEPs and metabolites related to HBTMD intervention. Network pharmacology, molecular docking, and molecular dynamics simulations explored potential molecular interactions and candidate targets. Ferrostatin-1 (Fer-1), Mdivi-1, and p53 siRNA were used in the indicated cell experiments to explore ferroptosis-associated injury and possible mitochondrial regulatory mechanisms. Results UHPLC-MS/MS identified 684 chemical constituents in HBTMD. Pharmacodynamic evaluation showed that HBTMD significantly reduced carotid lipid accumulation and plaque area in CAS mice. Integrated proteomic and metabolomic analyses suggested that modulation of autophagy and ferroptosis pathways may underlie HBTMD’s efficacy. Network pharmacology predicted p53 as a core target. Molecular docking and MD simulations suggested plausible interactions of spinosin and salvianolic acid A with p53, without establishing direct target engagement. In vitro and in vivo validation showed that HBTMD improved mitochondrial integrity, reduced inflammatory cytokines, and inhibited ferroptosis. HBTMD also lowered ROS and altered markers consistent with increased mitophagy-related activity; Mdivi-1 partially attenuated selected protective effects. The additional effects of HBTMD on several pathway-associated proteins were diminished after p53 knockdown, supporting the involvement of p53 but not establishing a complete causal pathway. Conclusion These preclinical findings suggest that HBTMD attenuates ferroptosis-associated endothelial injury and vascular inflammation in experimental CAS, accompanied by changes in p53-Parkin signaling and mitophagy-related markers. The pathway warrants further causal and translational validation.