Network pharmacology study of Cassiae Semen in treatment of hyperlipidemia and experimental validation in zebrafish
Objective:To investigate the mechanisms of Cassiae Semen in treating hyperlipidemia based on network pharmacology and be validated through zebrafish experiments.Methods:The active ingredients and related targets of Cassiae Semen were searched and screened in the Bioinformatics Analysis Tool for Molecular Mechanisms of Traditional Chinese Medicine(BATMAN-TCM)and the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).Hyperlipidemia-related target genes were obtained through searching GeneCards,DisGeNET and Online Mendelian Inheritance in man(OMIM).The common targets of drugs and diseases were obtained by Venny 2.1.0 platform.The Cassiae Semen-active ingredients-potential target network diagram was drawn by Cytoscape3.8.2,the protein-protein interaction(PPI)network diagram was constructed by String database,and the Cassiae Semen-hyperlipidemia common targets were subjected to Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis through the Database for Annotation,Visualization and Integrated Discovery(DAVID).According to the results of the network pharmacology study,aurantio-obtusin(AO)was hypothesized to be the effective acting component of Cassiae Semen in lowering blood lipids,and then the zebrafish experiments were carried out to verify based on above.The zebrafish hyperlipidemia model was induced by high-fat diet with egg yolk liquid,and was treated by AO after modeling,in order to verify the effect and mechanisms of AO in treating hyperlipidemia.Firstly,the toxicity experiment of AO on hyperlipidemic zebrafish was performed to determine the maximum administration concentration of AO.Subsequently,the effects of AO on total cholesterol(TC)and triglyceride(TG)contents and liver histomorphometry of hyperlipidemic zebrafish were observed.Quantitative real-time PCR was used to detect the expressions of core target mRNAs in hyperlipidemic zebrafish.Results:Thirteen active components of Cassiae Semen including AO were obtained.A total of 109 potential targets of Cassiae Semen acting on hyperlipidemia were obtained,including the core targets tumor necrosis factor(TNF),interleukin(IL)-1β,prostaglandin-endoperoxide synthase 2(PTGS2),Caspase 3(CASP3),peroxisome proliferator activated receptor-γ(PPAR-γ),etc.,which were mainly involved in IL-17 signaling pathway,TNF signaling pathway,as well as lipid and atherosclerosis-related pathways.The results of zebrafish validation experiments showed that,the significant lipid accumulation in zebrafish after modeling occurred,and AO could significantly reduce the TG and TC contents in hyperlipidemic zebrafish tissue(P＜0.05,P＜0.01),reduce the fat vacuole and lipid droplet in liver,and significantly down-regulate the expressions of core targets TNF-α and IL-1 β mRNA(P＜0.01).Conclusion:Based on network pharmacology,the core pathways and targets of Cassiae Semen in the treatment of hyperlipidemia are obtained,and the improvement effect of AO on hyperlipidemia is preliminarily revealed by zebrafish experiments,it may play the therapeutic effect through the core targets such as TNF-α,IL-1 β,etc.,in order to provide a reference for the later study of the clinical application of AO in hyperlipidemia.
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