吴氏泻心汤加减方通过抑制TLR4/NF-κB通路改善小鼠非酒精性脂肪性肝病的炎症反应OA

Non-alcoholic fatty liver disease(NAFLD)is the most prevalent chronic liver disease worldwide,characterized primarily by hepatic inflammation and dysregulated lipid metabolism.As a classic representative formula of the"bitter-purging method",Modified Wushi Xiexin Decoction has demonstrated definite clinical efficacy;however,its anti-inflammatory effects and underlying mechanisms in the treatment of NAFLD remain unclear.This study combines network pharmacology with animal experimental validation to evaluate the therapeutic effect of Modified Wushi Xiexin Decoction on NAFLD and reveal its potential mechanisms of action.Drug targets were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).NAFLD-related disease targets were retrieved from the Therapeutic Target Database(TTD),GeneCards,Online Mendelian Inheritance in Man(OMIM),Pharmacogenomics Knowledge Base(PharmGKB),and Comparative Toxicogenomics Database(CTD).R software was used to screen the intersection targets of the two sets.Core target analysis and network visualization were performed by using the Search Tool for the Retrieval of Interacting Genes/Proteins(STRING)database and Cytoscape 3.8.2 software.R software was applied for Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Core targets were analyzed based on the Gene Expression Omnibus(GEO)database,and receiver operating characteristic(ROC)curves were plotted to evaluate the diagnostic efficacy of the core targets.For animal experimental validation,a NAFLD mouse model was established by feeding a 60%high-fat diet combined with intraperitoneal injection of streptozotocin.Pathological improvements were assessed by hematoxylin-eosin(HE)staining and oil red O staining.Enzyme-linked immunosorbent assay(ELISA)was used to detect the level of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)in liver tissue.Immunohistochemistry(IHC)and Western blot(WB)were performed to measure the expression of Toll-like receptor 4(TLR4),nuclear factor-κB(NF-κB),and NOD-like receptor protein 3(NLRP3)proteins in mouse liver tissue.Network pharmacology identified 97 active components,108 intersection targets,and 10 core targets of Modified Wushi Xiexin Decoction.GO enrichment analysis indicated that anti-NAFLD effects were exerted by regulating biological processes such as oxidative stress and nutritional levels.KEGG enrichment analysis mainly involved signaling pathways,including TLR,NF-κB,and TNF.The GEO database indicates that there is a significant difference in TLR4 expression between healthy individuals and NAFLD patients(P<0.05),with the area under the ROC curve(AUC)being greater than 0.810,suggesting a high diagnostic efficacy.Animal experiments showed that Modified Wushi Xiexin Decoction could alleviate pathological damage in the liver tissue of NAFLD mice,reduce blood lipids,improve liver function,and downregulate the expression of inflammatory factors in liver tissue.IHC and WB confirmed that Modified Wushi Xiexin Decoction could downregulate the TLR4 expression,inhibit NF-κB activity,and suppress NLRP3 inflammasome activation.In conclusion,Modified Wushi Xiexin Decoction may alleviate inflammatory responses and improve liver function and blood lipids in NAFLD by regulating the TLR4/NF-κB signaling pathway and inhibiting NLRP3 inflammasome activation,thereby exerting therapeutic effects on NAFLD.