芜菁的降血脂活性评价及作用机制探讨

Evaluation of hypolipidemic activity of Brassica rapa and its mechanism

目的:评价芜菁的降血脂活性及基于网络药理学的机制。方法:首先通过体外胆酸盐结合实验和油酸诱导的HepG2脂肪变性细胞模型评价芜菁的降血脂活性。再通过文献检索与SwissTargetPrediction、SEA Search Server平台,筛选芜菁降血脂的主要有效成分与作用靶点,运用Cytoscape 3.7.2软件构建“化合物-靶点”网络图。在String数据库中构建交集靶点的PPI网络图,在DAVID数据库中进行GO和KEGG通路的富集分析。结果:体外降血脂实验发现,芜菁水提物对甘氨胆酸钠、牛磺胆酸钠和脱氧胆酸钠都有一定的结合能力,7 g/L时结合率分别为36.01%、28.93%、78.55%;在油酸诱导的HepG2脂肪变性细胞中,芜菁水提物不仅对脂肪堆积具有显著抑制效果(P<0.05),还能有效降低细胞内甘油三酯和总胆固醇水平(P<0.01)。对芜菁降血脂作用机制分析中,通过检索文献收集到符合类药五原则芜菁活性成分21个,预测到682个潜在药物靶点,其中与高血脂相关的靶点有55个;从构建的“化合物-靶点”网络图中可以得出,6-姜酮酚、6-姜烯酚、苄基-β-D-吡喃葡萄糖苷、苄基α-D-果糖呋喃糖苷和甘草素的度值较大,可能是芜菁降血脂的主要活性成分;PPI网络图和KEGG通路分析显示,芜菁通过调节VEGFA、IL6、EGFR、PPARG等关键靶点,干预淀粉和蔗糖代谢、半乳糖代谢、胰岛素抵抗、HIF-1、PI3K-Akt等信号通路发挥其降血脂活性。结论:芜菁水提物在体外具有很好的降血脂活性,并运用网络药理学初步揭示了其治疗高血脂的多成分-多靶点-多途径的作用机制,为深入研究芜菁的药效物质基础和作用机制提供理论依据。

Objective:To evaluate the hypolipidemic activity of Brassica rapa and to explore its mechanism by network pharmacology approach.Methods:The hypolipidemic activity of Brassica rapa aqueous extract(BRAE)was evaluated by bile salt-binding capacity and oleic acid-induced HepG2 steatosis cell model.The active compounds of Brassica rapa were collected from literature,and targets were predicted from SwissTargetPrediction and SEA Search Server platform.Cytoscape 3.7.2 soft-ware was used to construct“compound-target”network.Protein-protein interaction(PPI)network was constructed by String plat-form.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses were conducted based on DAVID database.Results:In vitro experiment showed that BRAE exhibited excellent bile salt-binding capacity,and the binding rates of sodium glycylcholate,sodium taurocholate and sodium deoxycholate were 36.01%,28.93%and 78.55%at 7 g/L,respectively.BRAE showed a significant hypolipidemic activity on steatosis cells,which can reduce the accumulation of lipid drop-lets and the levels of triglyceride(TG)and total cholesterol(TC)compared with the model group(P<0.05).A total of 21 active components of Brassica rapa and 682 potential targets were obtained,among which 55 targets were associated with hyperlipid-emia.The“compound-target”network showed that 6-paradol,6-shogaol,benzyl-beta-D-glucopyranoside,benzyl-alpha-D-fructo-furanoside and liquiritigenin were core components.PPI network and KEGG enrichment analysis found that Brassica rapa could treat hyperlipemia by regulating the core targets,such as VEGFA,IL6,EGFR and PPARG,and affected 36 signaling pathways(including starch and sucrose metabolism,galactose metabolism,insulin resistance,HIF-1 and PI3K-Akt).Conclusion:BRAE has excellent hypolipidemic activity in vitro,and preliminarily reveals the multi-component,multi-target,multi-path mechanism in the treatment of hyperlipidemia by network pharmacology approach,which provides a scientific foundation for further study.