基于网络药理学和实验验证探讨化痰活血解毒方治疗动脉粥样硬化的作用机制OA

Objective To investigate the potential mechanism of the Huatan Huoxue Jiedu Formula in treating atherosclerosis,integrating network pharmacology prediction with experimental validation.Methods Network pharmacology:Active compounds of the formula,their related targets,and atherosclerosis-associated targets were predicted and screened using the TCMSP database,literature databases,and the DisGeNET database.The Venny 2.1.0 platform was utilized to identify core targets of the formula against atherosclerosis.GO and KEGG enrichment analyses were performed using the Metascape database.A protein-protein interaction network was constructed using the STRING database."Drug-active ingredient-core target"and"signaling pathway-core target-active ingredient"networks were built using Cytoscape 3.8.2 software.Based on the analytical results,molecular docking validation was conducted.Animal experiment:Twenty ApoE-/-mice were randomly divided into a model group and a Huatan Huoxue Jiedu Formula group(n=10 per group).Both groups were fed a high-fat diet for 16 weeks.From the ninth week onwards,the formula group received the Huatan Huoxue Jiedu Formula by gavage for 8 weeks,while the model group received normal saline by gavage for 8 weeks.Ten C57BL/6J mice were selected as the normal group,fed a standard diet for 16 weeks,and received normal saline by gavage from the ninth week onwards.After the gavage period,all mice were euthanized for sample collection.En face Oil Red O staining was used to assess aortic lipid deposition.Hematoxylin and eosin staining was employed to measure carotid plaque area.Serum levels of TNF-α,IL-6,IL-1β,and AGEs were detected by ELISA.The protein expression levels of p-P65/P65 and RAGE in the carotid artery were determined by Western Blot.Results Network pharmacology screening identified 79 active compounds in the Huatan Huoxue Jiedu Formula,corresponding to 1 486 related targets.A total of 239 atherosclerosis-related targets were identified.The intersection yielded 70 core targets.GO and KEGG enrichment analyses revealed 45 cellular components,1 376 biological processes,88 molecular functions,and 239 KEGG signaling pathways.Molecular docking validation demonstrated that the binding energies of all 15"key component-key target"pairs were less than 0 kcal·mol-1,indicating strong affinity between the key components and their targets.Experimental validation showed that the Huatan Huoxue Jiedu Formula reduced carotid plaque area,decreased aortic lipid deposition,lowered serum levels of TNF-α,IL-6,IL-1β,and AGEs,and inhibited the protein expression of p-P65/P65 and RAGE,thereby exerting an anti-atherosclerotic effect.Conclusion The Huatan Huoxue Jiedu Formula exerts its anti-atherosclerotic effect by modulating inflammatory responses through the inhibition of the AGE-RAGE/NF-κB signaling pathway,involving a synergistic mechanism of multiple components and multiple targets.