基于网络药理学研究大补元煎防治AD的作用机制及AMPK/SIRT1信号通路验证OA北大核心CSTPCD
Based on network pharmacology to explore mechanism of Dabuyuanjian against AD and verification of AMPK/SIRT1 signaling pathway
目的:通过网络药理学和分子对接技术探究大补元煎防治阿尔茨海默病(AD)的作用机制,并利用动物实验验证所发现的分子机制.方法:网络药理学分析大补元煎抗AD的有效成分及作用靶点.利用AutoDock和PyMOL软件对药物的核心成分与核心蛋白进行分子对接验证.AD模型小鼠给予大补元煎治疗,并验证所发现的核心通路.结果:共筛选出80个有效活性成分和107个疾病作用靶点.大补元煎治疗AD的作用靶点有95个,其中核心靶点有35个.GO富集发现主要涉及程序性细胞死亡过程、细胞凋亡和信号转导调节等.KEGG信号通路富集发现主要涉及磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路、Wnt信号通路、腺苷酸活化蛋白激酶(AMPK)信号通路等.Morris水迷宫实验显示,大补元煎可减少AD小鼠的平台潜伏期,并增加小鼠的穿越平台次数和目标象限时间.免疫组化实验(IHC)结果显示,大补元煎可增加AD小鼠海马CA3区神经元核抗原(NeuN)标记的阳性细胞数.免疫荧光(IF)结果显示,大补元煎可抑制AD小鼠海马CA3区胶质纤维酸性蛋白(GFAP)、离子钙结合蛋白1(IBA1)的表达水平.Western blot实验显示,大补元煎可增加AD小鼠海马中磷酸化的腺苷酸活化蛋白激酶α(AMPKα)和沉默信息调节因子1(SIRT1)的表达水平.结论:本研究探讨了大补元煎抗AD的作用机制,并发现大补元煎可通过激活AMPK/SIRT1信号通路改善AD认知损伤、神经元丢失和神经炎症.
Objective:To explore the mechanism of Dabuyuanjian in Against alzheimer's disease(AD)through network phar-macology and molecular docking technology,and to verify the molecular mechanism discovered by animal experiments.Methods:Net-work pharmacology was used to analyze the active ingredients and targets of AD in the treatment of large supplementary yuan decoc-tion.The core components of the drug were verified by molecular docking with the core protein by using AutoDock and PyMOL soft-ware.AD model mice were treated with Dabuyuanjian,and the core pathways which discovered were verified.Results:A total of 80 active ingredients and 107 disease targets were screened out.Dabuyuanjian had 95 targets in the treatment of AD,of which 35 were core targets.GO enrichment found that it mainly involved in programmed cell death process,apoptosis process and signal transduction regulation,etc.KEGG signaling pathway enrichment found that it mainly involved PI3K/Akt signaling pathway,Wnt signaling pathway,AMPK signaling pathway,etc.Morris water maze experiment showed that Dabuyuanjian could reduce the escape latency of AD mice,and increase the number of crossing platform and time's target quadrant.Immunohistochemistry(IHC)showed that Dabuyuanjian could increase the number of positive labeled-NeuN cells in the hippocampal CA3 region of AD mice.Immunofluores-cence(IF)showed that Dabuyuanjian could inhibit the expression levels of(GFAP)and ionized calcium-binding protein 1(IBA1)in the hippocampal CA3 region of AD mice.Western blot experiments showed that Dabuyuanjian could increase the expression levels of phosphorylated adenylate-activated protein kinase α(AMPKα)and silent information regulator 1(SIRT1)in the hippocampus of AD mice.Conclusion:This study explores the mechanism of Dabuyuanjian against AD,and find that Dabuyuanjian can improve cognitive impairment,neuron loss and neuroinflammation via activating AMPK/SIRT1 signaling pathway of AD.
田梦杰;龙清华;曾楚华;刘道忠;王平;袁林
湖北民族大学,湿性疾病发生与干预湖北省重点实验室,恩施 445000湖北中医药大学,老年医学研究所,武汉 430065
中医学
大补元煎阿尔茨海默病网络药理学AMPK/SIRT1信号通路
DabuyuanjianAlzheimer's diseaseNetwork pharmacologyAMPK/SIRT1 signaling pathway
《中国免疫学杂志》 2024 (008)
1692-1700 / 9
国家自然科学基金(82060831);湖北省自然科学基金青年项目(2021CFB206);湖北民族大学高水平科研成果校内培育项目(PY21015).
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