基于网络药理学探讨栀黄止痛散治疗类风湿性关节炎作用机制及实验验证  

Network pharmacology-based study on mechanism of Zhi-Huang-Zhi-Tong powder in rheumatoid arthritis treatment

作　　者：田晓云 杨莹洁 郑婉婷 黄鸣清[1] 赵海誉[2] 南丽红[1] 陈剑钰[1] TIAN Xiao-yun;YANG Ying-jie;ZHENG Wan-ting;HUANG Ming-qing;ZHAO Hai-yu;NAN Li-hong;CHEN Jian-yu(Dept of Pharmacology,School of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Institute of Chinese Materia Medica,China Academy of Chinese Medicine Sciences,Beijing 100700,China)

机构地区：[1]福建中医药大学药学院,福建福州350122 [2]中国中医科学院中药研究所,北京100700

出　　处：《中国药理学通报》2024年第2期381-389,共9页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81903629);福建省自然科学基金资助项目(No 2022J01868);福建中医药大学人才启动项目(No X2019-004)。

摘　　要：目的结合网络药理学及体外细胞实验探索栀黄止痛散(Zhi-Huang-Zhi-Tong powder,ZHZTP)治疗类风湿性关节炎(rheumatoid arthritis,RA)的有效物质基础及相关作用机制。方法使用数据库检索并筛选得到ZHZTP活性成分靶点和疾病靶点;交集得到共同靶点;构建“药物-成分-靶点”关系网络图对交集基因进行GO和KEGG富集分析;并将核心成分与核心靶点进行分子对接。最后基于HUVEC细胞体外炎症模型,采用MTT法、平板划痕实验及蛋白质印迹等方法验证ZHZTP的药效及机制。结果ZHZTP中筛选出核心成分熊果酸和大黄素;治疗RA的重要靶点AKT1、IL-6和TNF;富集分析中,GO结果表明,ZHZTP对细胞分子功能、生物学功能及细胞组成均有影响,KEGG结果表明,ZHZTP的功能主要富集在PI3K-AKT、TNF和IL-17信号通路;分子对接结果显示,核心成分与核心靶点具有较好结合作用。体外结果表明,不同浓度的ZHZTP均可抑制HUVECs的炎性增殖和迁移,并且能够抑制PI3K,AKT及m-TOR蛋白表达水平。结论ZHZTP可能通过PI3K/AKT/m-TOR信号通路调控RA疾病发展进程,从而发挥抗RA作用,其主要有效物质基础为熊果酸和大黄素。Aim To discover the potential active compounds and possible mechanisms in rheumatoid arthritis(RA)treatment with Zhi-Huang-Zhi-Tong powder(ZHZTP)by using network pharmacology and in vitro study.Methods The active ingredient targets and disease targets of Zhihuang Zhitong Powder were searched and screened by database;they intersected to get a common target;and the“drug-component-target”relationship network diagram was constructed for GO and KEGG enrichment analysis of the overlapping genes;then the core components were docked with the core targets.Finally,based on the inflammation model of HUVECs in vitro,the efficacy and mechanism of Zhihuang Zhitong powder were verified by MTT method,plate scratch test and Western blot.Results Active compounds involved in RA treatment were screened in the present study,and the top two were ursolic acid and emodin,all playing crucial roles in RA treatment with ZHZTP.Additionally,the key target was AKTA,TNF and IL-6.GO and KEGG enrichment analysis revealed that ZHZTP regulated BP,MF and CC,and also focused on regulating AKTA,TNF and IL-6 signaling pathway.Molecular docking showed that interactions between key active compounds and key targets were stable.In vitro ZHZTP significantly inhibited cell viability and migration of TNF-α-stimulated HUVECs,and the involved mechanism may be associated with PI3K/AKT/m-TOR signaling.Conclusions The present study reveals that the potential active compounds of ZHZTP are ursolic acid and emodin,and moreover,the involved mechanisms of ZHZTP for RA treatment are associated with PI3K/AKT/m-TOR signaling.

关 键 词：栀黄止痛散 类风湿关节炎 网络药理学 活性成分 分子对接 

分 类 号：R-332[医药卫生] R287R319R593.22