基于网络药理学与体外实验探讨茯苓酸治疗心肌纤维化的作用机制  

Explore the mechanism of Pachymic acid in the treatment of myocardial fibrosis based on network pharmacology and in vitro experiments

作　　者：吴婉婉 魏科东 丁芮 任涵 赵婉竹 陈明[1] 周鹏[1,3] 王靓 黄金玲[1,3] WU Wanwan;WEI Kedong;DING Rui;REN Han;ZHAO Wanzhu;CHEN Ming;ZHOU Peng;WANG Liang;HUANG Jinling(School of Integrated Chinese Medicine and Western Medicine,Anhui University of Chinese Medicine,Hefei 230012,China;School of Chinese Medicine,Anhui University of Chinese Medicine,Hefei 230012,China;Anhui Provincial Key Laboratory of TCM Compounds,Hefei 230012,China)

机构地区：[1]安徽中医药大学中西医结合学院,安徽合肥230012 [2]安徽中医药大学中医学院,安徽合肥230012 [3]中药复方安徽省重点实验室,安徽合肥230012

出　　处：《海南医学院学报》2024年第10期747-761,共15页Journal of Hainan Medical University

基　　金：国家自然科学基金面上项目(81973844,81373533)。

摘　　要：目的:基于网络药理学与体外验证实验探讨茯苓酸治疗心肌纤维化(myocardial fibrosis,MF)的作用机制。方法:借助SwissTargetPrediction、GeneCards等数据库预测茯苓酸、氧化应激及MF靶点,取三者交集靶点于STRING数据库构建蛋白互作(PPI)网络,借助Cytoscape 3.7.2软件进行可视化分析,筛选出核心靶点,使用Metascape数据库进行GO和KEGG富集分析,预测茯苓酸治疗MF的作用机制,采用分子对接技术及大鼠心肌成纤维细胞(cardiac fibroblasts,CFs)实验加以验证。结果:预测发现,茯苓酸潜在靶点164个、氧化应激靶点3040个、MF靶点4441个,三者交集靶点84个,涉及Bcl-2、PTGS2、Bcl-2L1、MMP-2等9个核心靶点。GO分析发现,生物过程主要作用于抗氧化活性、对缺氧的反应。KEGG分析显示,茯苓酸治疗MF的主要信号通路为PI3K/Akt。分子对接结果显示,茯苓酸与Bcl-2、PTGS2、Bcl-2L1、MMP-2等9个核心靶点均具有良好的结合活性。体外实验结果显示,茯苓酸(5、10、20μmol/L)能显著抑制CFs迁移能力(P<0.01),降低ROS、MDA水平(P<0.05),升高SOD水平(P<0.05),下调CollagenⅠ、CollagenⅢ、MMP-9、MMP-2、PTGS2 mRNA水平(P<0.05)和CollagenⅠ、CollagenⅢ、α-SMA蛋白表达(P<0.05),上调PI3K、Akt、Bcl-2、Bcl-2L1 mRNA水平(P<0.05)和p-PI3K、p-Akt蛋白表达(P<0.05)。采用PI3K抑制剂(LY294002)后,茯苓酸对CFs细胞中ROS、SOD、MDA、CollagenⅠ、CollagenⅢ、α-SMA、p-PI3K、p-Akt作用被逆转(P<0.05)。结论:表明茯苓酸能够抑制CFs氧化应激损伤和心肌纤维化,该作用与其调控PI3K/Akt信号通路密切相关。Objective:To investigate the mechanism of Pachymic acid in the treatment of myocardial fibrosis(MF)based on network pharmacology and in vitro validation.Methods:The targets of Pachymic acid,oxidative stress and MF were predicted using SwissTargetPrediction,GeneCards and other databases,and the three intersection targets were selected to construct the protein interaction(PPI)network in STRING database.Visualization analysis was carried out by Cytoscape 3.7.2 software,and the core targets were selected.GO and KEGG enrichment analysis were performed using Metascape database to predict the mechanism of Pachymic acid in the treatment of MF,which was verified by molecular docking technique and rat cardiac fibroblasts(CFs).Results:According to the prediction,there were 164 potential targets of Pachymic acid,12139 oxidative stress targets,4441 MF targets and 84 intersection targets of the three,involving 9 core targets such as Bcl-2,PTGS2,Bcl-2L1 and MMP-2.GO analysis found that biological processes mainly act on antioxidant activity and response to hypoxia.KEGG analysis showed that the main signaling pathway of Pachymic acid in MF treatment was PI3K/Akt.The molecular docking results showed that Pachymic acid had good binding activity with 9 core targets such as Bcl-2,PTGS2,Bcl-2L1 and MMP-2.The results showed that Pachymic acid(5,10,20μmol/L)could significantly inhibit the migration of CFs(P<0.01),decrease the levels of ROS and MDA(P<0.05),and increase the level of SOD(P<0.05),down-regulated mRNA levels of CollagenⅠ,CollagenⅢ,MMP-9,MMP-2 and PTGS2(P<0.05)and protein expressions of CollagenⅠ,CollagenⅢandα-SMA(P<0.05).Pachymic acid also up-regulated mRNA levels of PI3K,Akt,Bcl-2 and Bcl-2L1(P<0.05),and protein expressions of p-PI3K and p-Akt(P<0.05).After the use of PI3K inhibitor(LY294002),the effects of Pachymic acid on ROS,SOD,MDA,CollagenⅠ,CollagenⅢ,α-SMA,p-PI3K and p-Akt in CFs cells were reversed(P<0.05).Conclusion:The results show that Pachymic acid can inhibit oxidative stress injury of CFs and myoc

关 键 词：茯苓酸 心肌纤维化 网络药理学 分子对接 

分 类 号：R285[医药卫生—中药学]