天然产物研究与开发 ›› 2023, Vol. 35 ›› Issue (3): 509-518.doi: 10.16333/j.1001-6880.2023.3.017

• 数据研究 • 上一篇    下一篇

基于网络药理学和实验验证探讨黄连解毒汤治疗血管性痴呆的作用机制

张运辉1,2,杨梦琳1,2*,伍大华2,3,刘   霞1,杨   昆1,李   祥1   

  1. 1重庆三峡医药高等专科学校,重庆 404120;2湖南中医药大学,长沙410208;3湖南省中医药研究院附属医院,长沙410006
  • 出版日期:2023-03-28 发布日期:2023-03-28
  • 基金资助:
    国家自然科学基金(81874462);重庆市中医药重点学科(中医基础理论)建设项目(渝中医〔2021〕16号);湖南省研究生创新课题(2020CX10)

Mechanism of Huanglian Jiedu Decoction in treating vascular dementia based on network pharmacology and experimental verification

ZHANG Yun-hui1,2,YANG Meng-lin1,2*,WU Da-hua2,3,LIU Xia1,YANG Kun1,LI Xiang1   

  1. 1Chongqing Three Gorges Medical College,Chongqing 404120,China;2Hunan University of Chinese Medicine,Changsha 410208,China;3Affiliated Hospital of Hunan Provincial Academy of Traditional Chinese Medicine,Changsha 410006,China
  • Online:2023-03-28 Published:2023-03-28

摘要:

运用网络药理学方法探讨黄连解毒汤治疗血管性痴呆(vascular dementia,VD)的作用机制。通过 TCMSP数据库检索黄连解毒汤的有效成分及相关靶点;采用DisGeNET和GeneCards数据库搜集VD靶点,通过String在线数据库构建靶蛋白相互作用网络,采用R语言对关键靶点进行GO和KEGG富集分析。采用谷氨酸诱导PC12细胞损伤作为VD细胞模型,通过MTT法检测细胞存活率,利用试剂盒检测超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、丙二醛(malondialdehyde,MDA)含量;最后采用ELISA法和Western blot实验对网络药理学主要预测的生物过程与信号通路进行验证。结果共获得黄连解毒汤63个有效成分,对应靶点216个,与VD相关靶点共105个,KEGG相关信号通路前20条,GO分析前20个生物学过程。细胞实验证实了黄连解毒汤能有效增加谷氨酸诱导PC12细胞的存活率,抑制其炎症反应和氧化应激反应,降低JNK、p-JNK、FoxO1、LC3Ⅱ/Ⅰ、Beclin1、Atg7、Caspase-3、Caspase-9蛋白表达,升高p-FoxO1、P62蛋白表达。黄连解毒汤可能是通过抑制JNK/FoxO1通路进而抑制炎症反应、氧化应激反应、自噬和细胞凋亡而发挥治疗VD作用。

关键词: 黄连解毒汤, 网络药理学, 血管性痴呆, 自噬

Abstract:

To study the mechanism of Huanglian Jiedu Decoction in the treatment of vascular dementia(VD) by network pharmacology.Searched the effective ingredients and related targets of Huanglian Jiedu Decoction by the TCMSP database,then obtained the targets of VD through the DisGeNET and GeneCards database.Constructed protein interaction network map by String online database,and to performed GO and KEGG enrichment analysis of key targets by R language.Induction of PC12 cell injury by adopted glutamate as VD cell model.Detection of cell viability by the MTT assay,and reactive superoxide dismutase (SOD),glutathione peroxidase(GSH-Px),malondialdehyde(MDA) content by using Kits.ELISA and Western blot were used to verify the main biological processes and signaling pathways as predicted by network pharmacology.A total of 63 active ingredients of Huanglian Jiedu Decoction were screened,216 corresponding targets,105 targets related to VD,the top 20 KEGG related signaling pathways and GO analysis of the top 20 biological processes.Cell experiments confirmed that Huanglian Jiedu Decoction effectively increased the survival rate of glutamate-induced PC12 cells,inhibited the inflammatory response and oxidative stress response,reduced the protein expression of JNK,p-JNK,FoxO1,LC3Ⅱ/Ⅰ,Beclin1,Atg7,Caspase-3,Caspase-9,increased the protein expression of p-FoxO1 and P62.Huanglian Jiedu Decoction had a therapeutic effect on VD by inhibited the JNK/FoxO1 pathway,thereby inhibited inflammation,oxidative stress,autophagy and apoptosis.

Key words: Huanglian Jiedu Decoction, network pharmacology, vascular dementia, autophagy

中图分类号:  R285