[Mechanism study of Zhizi Dahuang Decoction in treating atherosclerosis based on network pharmacology, molecular...

This paper investigated the effects and mechanisms of Zhizi Dahuang Decoction(ZZDHT) in treating atherosclerosis(AS) through a comprehensive approach combining network pharmacology, molecular docking, and in vivo experiments. Active components and target sites of ZZDHT were identified and screened using TCMSP, SwissTargetPrediction, and SEA databases. AS-related target sites were obtained from GeneCards, OMIM, and TTD databases. Network pharmacology was employed to predict the core chemical constituents, core target sites, and potential mechanisms of ZZDHT in treating AS. Molecular docking was employed to evaluate their binding activity between the core components and targets. Core chemical components identified in ZZDHT included quercetin,(+)-dehydrodiconiferyl alcohol, luteolin, and 3',4',5,7-tetramethoxyflavone. KEGG enrichment pathways primarily encompass core targets such as Toll-like receptor 4(TLR4), nuclear factor-κB(NF-κB), interleukin(IL)-6, IL-1β, and tumor necrosis factor(TNF). Molecular docking revealeda strong affinity between the core chemical components and these core targets. Forty male apolipoprotein E knockout(ApoE~(-/-)) mice were randomly assigned to a model group(MOD), low-dose ZZDHT group(ZZDH-L, 2.21 g·kg~(-1)), medium-dose ZZDHT group(ZZDH-M, 4.42 g·kg~(-1)), high-dose ZZDHT group(ZZDH-H, 8.84 g·kg~(-1)), and atorvastatin calcium group(ATO, 0.01 g·kg~(-1)). Additionally, C57BL/6J male mice served as the control group(CON), with 8 mice per group. CON was fed a normal diet, while the other groups were fed a high-fat diet to build an AS mouse model. All groups received corresponding drugs via oral gavage, and the CON and MOD administered an equal volume of 0.9% sodium chloride solution for 8 consecutive weeks. Experimental results showed that compared with MOD, ZZDHT intervention significantly inhibited aortic plaque formation in ApoE~(-/-) mice, reduced inflammatory infiltration at plaque sites, downregulated serum IL-6, IL-1β, and TNF-α levels while increasing IL-10 levels, thereby reshaping inflammatory homeostasis. Simultaneously, it regulated blood lipids by lowering serum total cholesterol(TC), triglycerides(TG), and low-density lipoprotein cholesterol(LDL-C) levels and increasing high-density lipoprotein cholesterol(HDL-C). qRT-PCR and Western blot confirmed that after treatment, the mRNA content of inducible nitric oxide synthase(iNOS) in aortic tissue decreased significantly, and the mRNA and protein expression of TLR4, myeloid differentiation primary response gene 88(MyD88), and NF-κB were all significantly reduced. Based on network pharmacology and experimental validation, ZZDHT may exert therapeutic effects on AS by modulating the TLR4/NF-κB signaling pathway to inhibit inflammatory responses, regulate blood lipids, reduce plaque lipid deposition, and enhance plaque stability.