基于网络药理学探究戊己丸对代谢性炎症的作用机制及实验验证

Wuji Pills(戊己丸)Treats Metabolic Inflammation:a Study Based on Network Pharmacology and Animal Experiment

目的:利用网络药理学和动物实验探究戊己丸治疗代谢性炎症的作用机制。方法:利用中药系统药理学分析平台(TCMSP)检索戊己丸中黄连、吴茱萸的化学成分和作用靶点;通过GeneCards、OMIM等数据库查询靶点对应的基因,Cytoscape 3.7.2构建戊己丸化合物-靶点网络,通过STRING数据库构建活性化合物作用靶点PPI网络,利用Bioconductor对作用靶点进行GO功能和KEGG通路富集分析,预测其作用通路,并通过动物实验验证戊己丸治疗代谢性炎症相关通路蛋白水平。结果:网络药理学筛选出主要活性成分有槲皮素、异鼠李素、植物固醇、氢化小檗碱、小檗碱,涉及IL6、CASP3、VEGFA、EGFR、MYC等关键靶点。涉及脂肪酸代谢过程、细胞凋亡等分子过程,细胞外基质、肌浆网等细胞组成,氧化还原等分子功能,涉及PI3K-AKT、MAPK、IL-17、NF-κB、FOXO、AMPK等信号通路。动物实验结果显示,与模型对照组相比,戊己丸3 g/kg组血清TC含量显著降低,肝脏组织炎性病变改善,p-PI3K、p-AKT、p-eNOS蛋白表达显著上调,肝脏中Il6、Il18、Tnfa基因表达明显下调,Il10基因表达明显上调(P<0.05或P<0.01)。结论:戊己丸中活性化合物能通过改善细胞凋亡及氧化还原反应调节炎症因子治疗代谢性炎症。

Objective:To explore the mechanism of Wuji Pills(戊己丸)in treating metabolic inflammation by network pharmacology and animal experiments.Methods:The chemical constituents and targets of Huanglian(黄连)and Wuzhuyu(吴茱萸)in Wuji Pills was retrieved from the Traditional Chinese Medicine Pharmacology Systems Database and Analysis Platform(TCMSP).The genes corresponding to the targets were obtained from GeneCards,Online Mendelian Inheritance in Man(OMIM).Cytoscape 3.7.2 was employed to establish the compound-target network of Wuji Pills,and STRING to establish the protein-protein interaction(PPI)network.Bioconductor was used for the gene ontology(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment of the targets.Animal experiments were carried out to study the changes in the expression levels of the proteins involved in the predicted pathways associated with the treatment of metabolic inflammation by Wuji Pills.Results:The main active constituents of Wuji Pills included quercetin,isorhamnetin,phytosterol,hydroberberine,and berberine,which corresponded to the targets such as IL6,CASP3,VEGFA,EGFR,and MYC.The GO annotation predicted the biological processes such as fatty acid metabolism and cell apoptosis,the cell components such as extracellular matrix and sarcoplasmic reticulum,and the molecular functions such as redox.The key targets were mainly enriched in PI3K-Akt,MAPK,IL-17,NF-kB,FOXO,and AMPK signaling pathways.The results of animal experiments showed that compared with the model group,Wuij Pills(3 g/kg)lowered the total cholesterol,reduced the inflammatory lesions of the liver tissue,up-regulated the protein levels of p-PI3K,p-AKT,and p-eNOS,down-regulated the mRNA levels of Il6,Il18 and Tnfα,and up-regulated the mRNA level of Il10(P<0.05 or P<0.01).Conclusion:The active compounds in Wuji Pills can treat metabolic inflammation by improving cell apoptosis and redox reaction and regulating inflammatory cytokines.