基于网络药理学及药理实验验证探讨仙鹤草抗肝纤维化的作用机制

Anti-liver Fibrosis Mechanism of Agrimoniae Herba Based on Network Pharmacology and Experimental Verification

目的:探究仙鹤草抗肝纤维化的作用机制。方法:根据多种规则筛选仙鹤草化学成分,预测获得靶基因,与肝纤维化发病机制相关的靶点进行比对获得仙鹤草抗肝纤维化的可能作用靶点。通过生物学信息注释库DAVID对关联靶点进行基因本体(GO)富集分析和京都基因和基因组百科全书(KEGG)富集分析,采用Cytoscape构建仙鹤草化合物-靶点-信号通路相互作用网络图。进一步采用实验验证仙鹤草对大鼠肝星状细胞-T6(HSC-T6)肝纤维化指标α-平滑肌肌动蛋白(α-SMA)、胶原1型α1(COL1A1)及蛋白激酶B(AKT)蛋白磷酸化的影响。结果:关键靶点主要为AKT1、STAT3、Caspase-3、Jun、ESR1等,主要参与调控肿瘤信号通路、PI3K/AKT信号通路、肿瘤蛋白聚糖、胰岛素抵抗和FoxO信号通路等。仙鹤草提取物可抑制HSC-T6中α-SMA、COL1A1蛋白的表达,升高AKT的磷酸化水平,验证了网络药理学分析的部分预测结果。结论:仙鹤草可通过作用于PI3K/AKT信号通路等发挥抗肝纤维化作用。

Objective:To explore the anti-liver fibrosis mechanism of Agrimoniae Herba.Methods:The chemical compounds of Agrimoniae Herba were screened out by multiple methods and the target genes were predicted and obtained.The potential targets of Agrimoniae Herba against liver fibrosis were obtained through the comparison with the targets related to the pathogenesis of liver fibrosis.Gene Ontology(GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis were carried out for associated targets through DAVID.The chemical compound-target-pathway interaction network was visualized by Cytoscape software.The effects of Agrimoniae Herba on the expression of α-smooth muscle actin(α-SMA) and collagen type 1 alpha 1(COL1A1) and the phosphorylation of protein kinase B(AKT) in rat hepatic stellate cell-T6(HSC-T6) were further verified by experiments.Results:The key targets were mainly AKT1,STAT3,caspase-3,Jun, and ESR1,which were mainly involved in the regulation of tumor signaling pathway, PI3K/AKT signaling pathway, tumor proteoglycans, insulin resistance, FoxO signaling pathway, etc.Agrimoniae Herba extract could inhibit the protein expression of α-SMA and COL1A1 and increase the phosphorylation level of AKT in HSC-T6,which verified the partial prediction results of network pharmacological analysis.Conclusion:Agrimoniae Herba could inhibit liver fibrosis by acting on the PI3K/AKT and other signaling pathways.