目的 探讨中风醒脑液(Zhongfeng Xingnao Liquid)对脑出血(intracerebral hemorrhage,ICH)后神经系统及免疫系统的影响.方法 从中医药系统药理数据库和分析平台(Traditional Chinese Medicine Database and Analysis Platform,TCMSP)获取中风醒脑液有效活性成分;并通过TCMSP、SwissTargetPrediction数据库获取有效活性成分作用靶点;通过药物靶标在线数据库(Therapeutic Target Database,TTD)、人类在线孟德尔遗传数据库(Online Mendelian Inheritance in Man,OMIM)、DisGeNET、GeneCards和Drugbank数据库筛选脑出血相关靶点;通过京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)筛选神经系统和免疫系统相关靶点;运用STRING平台分别对"中风醒脑液-脑出血-免疫系统","中风醒脑液-脑出血-神经系统"交集靶点进行蛋白互作分析和筛选信号通路,运用MCC法筛选中风醒脑液影响脑出血后神经系统和免疫系统的核心靶点;利用Cytoscape 3.9.0 软件构建"药物-成分-核心靶点-通路"网络,计算网络度值(Degree),筛选出中风醒脑液关键活性成分,运用Autodock Vina软件对活性成分与核心靶点进行分子对接.利用胶原酶Ⅶ型诱导大鼠脑出血疾病模型进行实验验证.结果 预测得出中风醒脑液主要活性成分 31 种;中风醒脑液对脑出血后神经系统潜在作用靶点 33 个,对免疫系统潜在作用靶点 68 个;槲皮素、洋川芎醌和β-谷甾醇等是中风醒脑液影响脑出血后神经系统和免疫系统的主要活性成分,预测得出肿瘤蛋白P53(TP53)、丝裂原活化蛋白激酶 1(MAPK1)、胱天蛋白酶 3(CASP3)等是中风醒脑液影响脑出血后神经系统的核心靶点,肿瘤坏死因子(TNF)、白细胞介素 6(IL-6)、基质金属蛋白酶 9(MMP9)等是中风醒脑液影响脑出血后免疫系统的核心靶点;分子对接结果显示MMP9、信号转导及转录激活蛋白 3(STAT3)和白细胞介素 1β(IL-1β)等蛋白受体与槲皮素,洋川芎醌,β-谷甾醇及人参皂苷Rh2 等主要活性成分具有较好的亲和力.动物实验验证结果表明,中风醒脑液高剂量组能改善脑出血大鼠神经缺损症状及血肿周边组织病理结构,有效减少脑出血后CASP3、IL-1β、IL-6、TNF、PRKCA、PRKCD、RELA、TP53和MAPK1 mRNA的相对表达,上调NFKBIA、AKT1 mRNA的相对表达(P＜0.05,P＜0.01,P＜0.001).结论 中风醒脑液可能通过调控TNF、IL-6、TP53、MAPK1 及CASP3 等表达来影响脑出血后神经系统和免疫系统,发挥治疗作用.
Exploration of the Influence of Zhongfeng Xingnao Liquid on the Treatment of Nervous System and Immune System After Intracerebral Hemorrhage Based on Bioinformatics Analysis
Objective To explore the influence of Zhongfeng Xingnao Liquid(ZFXN)on nervous system and immune system after intracerebral hemorrhage(ICH).Methods The active components of ZFXN from Traditional Chinese Medicine Database and Analysis Platform(TCMSP)were screened.The targets of the active components were predicted by TCMSP and SwissTargetPrediction databases.The targets related to intracerebral hemorrhage were screened by using Therapeutic Target Database(TTD),Online Mendelian Inheritance in Man(OMIM),DisGeNET,GeneCards and Drugbank databases.Then,the targets related to nervous system and immune system were screened by using the Kyoto Encyclopedia of Genes and Genomes(KEGG).The common targets of"ZFXN-intracerebral hemorrhage-immune system"and"ZFXN-intracerebral hemorrhage-nervous system"were imported into STRING platform to conduct protein-protein interaction(PPI)network analysis and signaling pathways screening,respectively.The key targets of the influence of ZFXN on nervous system and immune system after ICH were screened by MCC method.The"drugs-components-key targets-pathways"network was constructed by Cytoscape 3.9.0.software,and key active components in ZFXN were predicted by the degree of network.Autodock Vina software was used to investigate the molecular docking between the key active components and the key targets.Rat model of collagenase Ⅶ-induced intracerebral hemorrhage was used to verify the simulated results.Results A total of 31 active components,68 common targets related to immune system,and 33 common targets related to nervous system were obtained.Quercetin,β-sitosterol,and senkyunone are the main active components of ZFXN,which affect immune and nervous system after intracerebral hemorrhage.TP53,MAPK1,and CASP3 are the key targets of ZFXN affecting the nervous system.TNF,IL-6 and MMP9 are the key targets of ZFXN affecting the immune system.The results of molecular docking showed that quercetin,senkyunone,β-sitosterol,ginsenoside Rh2 and other key components have good binding activity with the key targets,such as MMP9,STAT3,IL-1β,etc..The results of animal experiments suggested that ZFXN could improve the neurological function and the pathological structure of the tissues around hematoma in rats with intracerebral hemorrhage,effectively reduce the mRNA relative expressions of CASP3,IL-1β,IL-6,TNF,PRKCA,PRKCD,RELA,TP53 and MAPK1 after intracerebral hemorrhage,and increase the mRNA relative expressions of NFKBIA and Akt1(P＜0.05,P＜0.01,P＜0.001).Conclusion ZFXN may regulate the expressions of TNF,IL-6,TP53,MAPK1,CASP3 to intervene nervous system and immune system after intracerebral hemorrhage and to play a therapeutic role.
NETL
NSTL
万方数据
