基于网络药理学和实验验证探讨赤芍治疗胆汁淤积性肝炎的分子机制

Study on the Effective Components and Molecular Mechanismof Radix Paeoniae Rubra in the Treatment of Cholestatic Liver Disease based on Network Pharmacology

为探讨赤芍(Paeoniae Rubra Radix,PRR)治疗胆汁淤积性肝炎(Cholestatic Hepatitis,CH)的分子机制,本研究从TCMSP数据库获取PRR的化学成分,口服生物利用度OB≥30%,类药性DL≥0.18,筛选PRR潜在活性成分,通过PharmMapper和Swiss Target Prediction数据库预测PRR化学成分的作用靶点,通过Drugbank、TTD和Genecards数据库获取CH的相关靶点,利用在线平台Venny 2.1.0获得PRR化学成分和CH的共同靶点,利用STRING数据库构建蛋白-蛋白相互作用(PPI)网络,利用Cytoscape 3.9.0进行PPI网络可视化,通过DAVID数据库进行GO功能富集分析及KEGG通路富集分析,利用Cytoscape 3.9.0软件构建“成分-靶点-通路”,采用ANIT建立大鼠胆汁淤积模型,给予赤芍提取物(PRRE)进行干预,通过免疫组化技术考察各组大鼠肝脏组织蛋白表达情况。通过网络药理学分析筛选得到PRR的潜在活性成分28个,PRR潜在活性成分治疗CH的潜在靶点34个,并根据degree值筛选出TNF-α、EGFR、VEGFA等6个核心靶点,KEGG通路富集分析PRR通过MAPK等信号通路发挥治疗CH作用,免疫组化试验结果显示,与模型组比较,PPR给药组TNF-α、EGFR、VEGFA的表达均显著降低,差异具有统计学意义(P<0.05)。PRR可能通过芍药新苷、芍药苷干预TNF-α、EGFR、VEGFA等靶点进而调节MAPK等信号通路发挥治疗CH的作用。

To investigate the molecular mechanism of Paeoniae Rubra Radix(PRR)in the treatment of Cholestatic Hepatitis(CH).The chemical components of PRR were retrieved by TCMSP database and the underlying active ingredients was screened by the OB and DL.The potential targets of active ingredients of PRR were predicted by Pharmmapper and Swiss Target Prediction database.The related targets of CH were obtained by DrugBank,TTD and Genecards database.The common targets of active ingredients and CH was acquired by using Venny2.1.0 online platform.PPI network was constructed through STRING database and was visualized by Cytoscape 3.9.0.GO function enrichment analysis and KEGG pathway enrichment analysis were carried out through DAVID database.‘Components-targets-pathways’network was constructed through Cytoscape 3.9.0 software.ANIT was used to establish rat model of cholestasis,and Paeoniae Rubra Radix extract(PRRE)was administered for intervention,the protein expression in the liver tissue of the rats in each group was investigated by immuno-histochemistry(IHC).28 potential active ingredients.34 intersection targets of PRR in the treatment of CH were screened by Venny2.1.0 online platform.6 key targets were screened according to the degree value.The results of IHC showed that the expressions of EGFR,VEGFA,and p38MAPK in the PPRE administration group were significantly decreased compared with the model group,and the difference was statistically significant(P<0.05).PRR may play a role in the treatment of CH by intervening EGFR,VEGFA,p38MAPK and other targets through paeoniflorin and paeoniflorin,thereby regulating MAPK and other signaling pathways.