基于网络药理学探讨三黄泻心汤治疗脑出血的作用机制  

Mechanism of Sanhuang Xiexin Decoction in the Treatment of Intracerebral Hemorrhage Based on Network Pharmacology

作　　者：官玥玥 罗晓琼 黄利 余鑫源 李陈渝[1] 舒建中[1] 唐军[1] GUAN Yue-yue;LUO Xiao-qiong;HUANG Li;YU Xin-yuan;LI Chen-yu;SHU Jian-zhong;TANG Jun(Chongqing Hospital of Traditional Chinese Medicine,Chongqing 400021)

机构地区：[1]重庆市中医院,重庆400021

出　　处：《世界中西医结合杂志》2022年第11期2190-2199,2205,共11页World Journal of Integrated Traditional and Western Medicine

基　　金：国家中医药管理局中医药循证能力建设项目(2019XZZX-NB014);“重庆英才计划”项目(渝委人才[2021]5号);重庆市自然科学基金面上项目(cstc2020jcyj-msxmX0985)。

摘　　要：目的基于网络药理学探讨三黄泻心汤治疗脑出血(Intracerebral hemorrhage,ICH)的作用靶点及机制。方法借助TCMSP数据库、OMIM数据库、TTD数据库、GeneCards数据库,对三黄泻心汤作用于ICH的靶点进行预测,利用STRING数据库进行蛋白相互作用分析,采用Cytoscape软件对PPI网络进行优化及核心靶点的网络拓扑学分析。再通过Metascape数据库对交集靶点进行GO与KEGG富集分析探索其可能的分子作用机制。结合GO与KEGG富集分析结果,选择PI3K/AKT信号通路进行进一步的动物实验验证。通过注射IV型胶原酶的方法建立ICH大鼠模型,分为假手术组、模型组、三黄泻心汤低剂量组、三黄泻心汤高剂量组,连续干预14d后检测各组大鼠的mNSS评分,运用Western Blot检测各组大鼠脑组织中PI3K、p-PI3K、AKT、p-AKT蛋白表达水平,qPCR检测PI3K、AKT的mRNA表达水平。结果网络药理学筛选出,三黄泻心汤治疗ICH核心靶点22个,包括AKT1、IL6、TNF、VEGFA等,KEGG富集分析预测三黄泻心汤主要是通过HIF-1信号通路、IL-17信号通路、PI3K/AKT信号通路等发挥治疗作用。动物实验显示,与模型组比较,三黄泻心汤低、高剂量组mNSS评分均明显降低;三黄泻心汤高剂量组中PI3K、p-PI3K、AKT、p-AKT以及三黄泻心汤低剂量组中AKT、p-AKT蛋白表达明显升高;三黄泻心汤高剂量组和低剂量组中PI3K、AKT的mRNA表达水平均明显升高。结论三黄泻心汤治疗ICH具有多成分、多靶点、多通路的特点,可通过激活PI3K/AKT信号通路发挥治疗作用。Objective To explore the targets of Sanhuang Xiexin Decoction on the treatment of intracerebral hemorrhage(ICH)and its mechanism based on network pharmacology.Methods With the help of Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)database,Online Mendelian Inheritance in Man(OMIM)database,Therapeutic Target Database,and Gene Cards database,this paper predicted the targets of Sanhuang Xiexin Decoction on ICH and used the STRING database for protein-protein interaction(PPI)analysis.Cytoscape was used to optimize the PPI network and conduct network topology analysis for core targets.Then,Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed on the intersection targets through the Metascape database to explore the possible molecular mechanisms.Combined with the results of GO functional enrichment analysis and KEGG pathway enrichment analysis,the phosphoinositide 3 kinase/protein kinase B(PI3 K/AKT)signaling pathway was selected for further animal experiments.The rat model of ICH was induced by injecting type IV collagenase,and the rats were divided into a sham operation group,a model group,a low-dose Sanhuang Xiexin Decoction group,and a high-dose Sanhuang Xiexin Decoction group.After continuous intervention for 14 days,the mNSS scores of rats in each group were tested.Western blotting assay was used to determine the protein expression levels of PI3 K,p-PI3 K,AKT,and p-AKT in the brain tissues of rats in each group,and real-time quantitative polymerase chain reaction(RT-qPCR)was used to determine the mRNA expression levels of PI3 K and AKT.Results Twenty-twocore targets of Sanhuang Xiexin Decoction in the treatment of ICH were screened out by network pharmacology,including AKT1,interleukin(IL)-6,tumor necrosis factor(TNF),and vascular endothelial growth factor-A(VEGFA).KEGG pathway enrichment analysis predicted that Sanhuang Xiexin Decoction exerted a therapeutic effect mainly through the hypo

关 键 词：三黄泻心汤 网络药理学 脑出血 PI3K/AKT信号通路 作用机制 

分 类 号：R285.6[医药卫生—中药学]