笔管草醇提物防治高脂血症的潜在药效物质及机制研究OA

Objective:To identify the core active constituents of the ethanol extract of(Ethanol extract of Equisetum debile Roxb.,EED)responsible for its therapeutic effects against hyperlipidemia,and to elucidate its mechanism of action via the"constituents-targets-pathways"axis,thereby providing a scientific basis for its clinical application and new drug development.Methods:An in vitro model of lipid accumulation was established using oleic acid-induced HepG2 cells.The lipid-lowering efficacy of EED was verified via Oil Red O staining.Systemic identification of the absorbed constituents and metabolites of EED in rat serum following oral administration was performed using UPLC-Q-TOF-MS/MS,assisted by online databases including PubChem and TCMSP.Potential targets of these serum constituents related to hyperlipidemia were screened using the Swiss Target Prediction database.A protein-protein interaction(PPI)network was constructed using the STRING platform,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis was conducted via the DAVID database.Molecular docking was performed using AutoDock Vina,and the binding modes and affinities between potential active constituents and key targets were visualized and analyzed with PyMOL.To further verify the regulatory effect of EED on the expression of key genes,the RT-qPCR technique was used to detect the changes in the mRNA levels of genes such as STAT3,HSP90AA1,EGFR,PTGS2,and ESR1 in HepG2 cells before and after drug administration.Results:EED concentration-dependently reduced both intracellular TG content and lipid droplet accumulation in oleic acid-induced HepG2 cells(P<0.05).Seventeen major absorbed constituents,primarily flavonoids such as naringenin and kaempferol,were identified in rat serum.Network pharmacology analysis revealed that 31 core hyperlipidemia-related targets,including STAT3,AKT1,EGFR,and HSP90AA1,were commonly modulated by key serum constituents like naringenin,kaempferol,acacetin,and quercetin.KEGG enrichment analysis indicated that these targets were significantly enriched in 15 core pathways,predominantly the Lipid and Atherosclerosis signaling pathway.Molecular docking confirmed strong binding affinities,with binding free energies below-9.0 kcal/mol for the complexes of PTGS2 with acacetin,PTGS2 with kaempferol,PTGS2 with liquiritigenin,and ESR1 with liquiritigenin,indicating stable binding modes.RT-qPCR detection results showed that the mRNA expression levels of STAT3,HSP90AA1,EGFR,and PTGS2 genes were significantly increased in the model group,while the expression of the ESR1 gene was significantly decreased.After EED intervention,the expression changes of these genes were all reversed.Conclusion:EED exerts its anti-hyperlipidemic effect through the synergistic interactions of a"flavonoid cluster-core targets-lipid metabolism-related pathways"network.The core pharmacodynamic substances are flavonoids,including liquiritigenin,kaempferol,and acacetin,with the PTGS2 and ESR1 signaling pathways potentially serving as critical mechanistic routes.