基于网络药理学和实验验证探讨毛菊苣治疗肝纤维化作用机制

Exploring the Mechanism of Cichorium Glandulosum in the Treatment of Liver Fibrosis Based on Network Pharmacology and Experimental Validation

关键词:  毛菊苣  肝纤维化  网络药理学  分子对接  实验验证

Key words:  Cichorium glandulosum  liver fibrosis  network pharmacology  molecular docking  experimental validation

摘要:

目的 采用网络药理学和实验验证的方法探究毛菊苣抗肝纤维化的作用机制。方法 借助TCMSP、Pubchem、SwissTargetPrediction、Genecards等数据库构建“成分-靶点-通路”作用网，STRING数据库预测毛菊苣抗肝纤维化作用靶点，应用DAVID数据库进行KEGG和GO富集分析，AUTODOCK对活性成分和关键靶点进行分子对接。用PDGF-BB诱导活化细胞，验证秦皮甲素、秦皮乙素、菊苣酸、咖啡酸、绿原酸和异绿原酸等6个化合物对HSC-T6细胞的增殖、凋亡以及对肝纤维化指标的影响，Western blotting检测各成分对HSC-T6细胞中Ras、ERK1、ERK2、C-fos、JNK蛋白表达。结果 网络药理学筛选到成分与肝纤维化的共同靶点239个，PPI分析显示SRC、STAT3、HSP90AA1等靶点为关键靶点，KEGG分析结果显示毛菊苣影响的通路包括癌症通路、代谢通路等，GO分析预测毛菊苣主要影响信号转导等过程，分子对接结果显示与成分能够良好结合的靶点蛋白为MAPK1，与蛋白能够良好结合的成分为秦皮乙素、咖啡酸和绿原酸。6种化合物对PDGF-BB诱导HSC-T6细胞活化增殖的抑制作用均强于模型组，且均具有逆转肝纤维化指标的作用，其中秦皮乙素活性最强(P<0.01)。与模型组相比，6种化合物干预后HSC-T6细胞中的Ras、ERK1、ERK2、C-fos、JNK表达量均有所下降。结论 毛菊苣中各成分均有不同的抗肝纤维化作用，其作用与抑制ERK/RAS通路激活的有关。

Abstract:

OBJECTIVE To investigate the mechanism of Cichorium glandulosum in the treatment of liver fibrosis by using network pharmacology and experimental validation. METHODS A "component-target-pathway" network was constructed with the help of TCMSP, Pubchem, SwissTargetPrediction and Genecards databases, and the STRING database was used to predict the targets of Cichorium glandulosum against liver fibrosis. KEGG and GO enrichment analysis was performed in the DAVID database, and molecular docking of active ingredients and key targets was docked in AUTODOCK. PDGF-BB was used to induce activation of cells and verify the effects of six compounds, including quercetin, quercetin, chicoric acid, caffeic acid, chlorogenic acid, and isochlorogenic acid, on the proliferation, apoptosis, and liver fibrosis indicators of HSC-T6 cells. Western blotting was used to detect the expression of Ras, ERK1, ERK2, C-fos, and JNK proteins in HSC-T6 cells. RESULTS Network pharmacology screened 239 common targets between the components and liver fibrosis, PPI analysis showed that SRC, STAT3, HSP90AA1 and other targets were key targets, KEGG analysis showed that the pathways affected by Cichorium glandulosum included cancer pathways, metabolic pathways, etc. GO analysis predicted that Cichorium glandulosum mainly affected processes such as signal transduction. The molecular docking results showed that the target that could bind well with the components MAPK1, and the components that could bind well with the target aesculetin, caffeic acid and chlorogenic acid. Compared with the model group, the inhibition effect of the six compounds on PDGF-BB-induced HSC-T6 cell activation was stronger, and all 6 compounds had the effects to reverse the index of liver fibrosis, in which aesculetin had the strongest activity(P<0.01). The expression of Ras, ERK1, ERK2, C-fos, and JNK in HSC-T6 cells decreased after the interventions of 6 compounds. CONCLUSION Each component of Cichorium glandulosum has different anti liver fibrosis effects, which are related to the inhibition of ERK/RAS pathway activation.