吉林大学学报(医学版) ›› 2023, Vol. 49 ›› Issue (2): 261-271.doi: 10.13481/j.1671-587X.20230201

• 基础研究 •    下一篇

温补脾肾经验方治疗视神经脊髓炎谱系疾病作用机制的网络药理学分析

宫玉霜1,付业繁2,许会静2,郭健2,相琳1,胡睿1,樊睿1,王捷3,李淼1(),孙美艳2()   

  1. 1.吉林大学中日联谊医院神经外科,吉林 长春 130033
    2.吉林医药学院检验学院实验中心,吉林 吉林 132013
    3.吉林大学中日联谊医院神经内科,吉林 长春 130033
  • 收稿日期:2022-05-12 出版日期:2023-03-28 发布日期:2023-04-24
  • 通讯作者: 李淼,孙美艳 E-mail:limiao@jlu.edu.cn;sunmy990@163.com
  • 作者简介:宫玉霜(1995-),女,山东省威海市人,在读硕士研究生,主要从事视神经脊髓炎谱系疾病的发病机制和治疗等方面的研究。
  • 基金资助:
    国家自然科学基金项目(81771304);吉林省科技厅科技发展计划项目(20200201623JC);吉林省卫健委卫生科研人才专项项目(2019SCZ037);吉林省教育厅科学技术研究项目(JJKH20210500KJ)

Network pharmacology analysis on mechanism of Tonifying Slpeen and Kidney Empirical Prescription in treatment of neuromyelitis optica spectrum disorders

Yushuang GONG1,Yefan FU2,Huijing XU2,Jian GUO2,Lin XIANG1,Rui HU1,Rui FAN1,Jie WANG3,Miao LI1(),Meiyan SUN2()   

  1. 1.Department of Neurosurgery,China-Japan Union Hospital,Jilin University,Changchun 130033,China
    2.Experiment Center,College of Laboratory Medicine,Jilin Medical University,Jilin 132013,China
    3.Department of Neurology,China-Japan Union Hospital,Jilin University,Changchun 130033,China
  • Received:2022-05-12 Online:2023-03-28 Published:2023-04-24
  • Contact: Miao LI,Meiyan SUN E-mail:limiao@jlu.edu.cn;sunmy990@163.com

摘要:

目的 通过网络药理学分析和动物实验探讨温补脾肾经验方(TSKEP)治疗视神经脊髓炎谱系疾病(NMOSD)的潜在作用靶点和信号通路,为治疗NMOSD提供理论依据。 方法 采用中药系统药理学数据库及分析平台(TCMSP)筛选TSKEP的化合物和靶点,通过GendCard等数据库确定NMOSD的相关靶点基因;构建化合物-靶点网络确定核心化合物;利用STRING数据库和Cytoscape 3.6.0软件中的Network Analyzer插件获得蛋白-蛋白互作(PPI)网络的拓扑参数并识别核心靶点;利用OmicShare平台对核心靶点进行基因本体论(GO)功能注释分析和京都基因与基因组百科全书(KEGG)通路富集分析并构建靶点-通路网络,以识别关键的生物过程和信号通路;基于AutoDock软件对核心化合物和核心靶点进行分子对接。18只小鼠分为对照组、NMOSD组和TSKEP组,每组6只,HE染色观察3组小鼠大脑组织病理形态表现,采用酶联免疫吸附试验(ELISA)法检测小鼠血清中相关细胞因子水平。 结果 动物实验,TSKEP组小鼠神经功能缺损评分与NMOSD组比较明显降低(P<0.01),同时TSKEP组小鼠中枢神经系统炎性细胞浸润程度较轻。共筛选出304个活性化合物和655个药物靶点,其中与TSKEP治疗NMOSD相关的靶点有43个。槲皮素、木犀草素和豆甾醇等可能是核心化合物,白细胞介素6(IL-6)、白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)等可能是潜在的核心靶点。KEGG通路富集分析及靶点-通路网络提示TSKEP主要通过14条信号通路发挥治疗NMOSD作用。分子对接验证结果表明核心化合物与核心靶点之间结合良好。与NMOSD组和对照组比较,TSKEP组小鼠血清中相关炎性细胞因子水平降低(P<0.05或P<0.01)。 结论 基于网络药理学和动物实验验证,TSKEP可通过多靶点和多条信号通路调控血清中炎性细胞因子的表达并促进神经功能的修复再生,减轻中枢神经系统的炎症反应,从而发挥治疗作用。

关键词: 视神经脊髓炎谱系疾病, 温补脾肾经验方, 网络药理学, 分子对接

Abstract:

Objective To discuss the potential targets and signaling pathways of Tonifying Slpeen and Kidney Empirical Prescription (TSKEP) in the treatment of neuromyelitis optica spectrum disorders(NMOSD) through network pharmacology analysis and animal experiment,and to provide the theoretical basis for the treatment of NMOSD. Method Traditional Chinese Medicine System Analysis Platform(TCMSP) Database was used to screen the chemical compounds and targets of TSKEP; the related target genes in NMOSD were comfirmed through GendCard and other Databases;the core compounds were obtained by building the compound-target network; the STRING Database and the Network Analyzer plugin in Cytoscape 3.6.0 software were used to obtain the topology parameters of protein-protein interaction (PPI) network and to identify the core targets; Omicshare platform was used to perform the Geno Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis and the target-pathway network was conducted to identify core key biological processes and signaling pathways; the molecular docking of core compounds and core targets was performed based on AutoDock software. Eighteen mice were divided into control group, NMOSD group, and TSKEP group, and there were 6 mice in each group. The morphology of brain tissue of the mice in various groups were observed by HE staining,and the levels of related cytokines in serum of the mice were detected by enzyme-linked immunosorbent assay(ELISA) method. Results The animal experiment results showed that compared with NMOSD group,the score of neural function defect of the mice in TSKEP group was decreased(P<0.01), and the degree of inflammatory cell infiltration in central nervous system of the mice in TSKEP group was allevited. A total of 304 active compounds and 655 drug targets were screened, and 43 potential targets were related to the treatment of NMOSD with TSKEP. Quercetin, luteolin, and stigmasterol may be the core compounds,and interleukin-6(IL-6),interlleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α) could be the poetential core targets.The KEGG pathway enrichment analysis and network analysis results showed that TSKEP may play an important role in the treatment of NMOSD through 14 signaling pathways.The molecular docking verification results showed that core compounds combined well with the core targets. Compared with NMOSD group and control group, the levels of serum inflammatory cytokines of the mice in TSKEP group were decreased(P<0.05 or P<0.01). Conclusion Based on the verification of network pharmacology and animal experiment, TSKEP can exert the therapeutic effect through regulating the expressions of serum inflammatory cytokines, promoting the repairment and regeneration of nerve function,and reducing the inflammatory response of the central nervous system by multiple targets and signaling pathways.

Key words: Neuromyelitis optica spectrum disorders, Tonifying Slpeen and Kidney Empirical Prescription, Network pharmacology, Molecular docking

中图分类号: 

  • R744.52