摘要
目的:通过网络药理学方法筛选交泰丸的有效化学成分和作用靶点,结合动物实验验证,探讨其治疗阿尔茨海默病(Alzheimer’s disease,AD)的作用机制。方法:本研究利用中药系统药理数据(TCMSP)检索交泰丸中黄连和肉桂2味中药的有效化学成分,经过口服生物利用度(OB)和类药性(DL)的筛选和相关文献的检索,得到药物的有效化学成分,并进行靶点的预测。通过GeneCards和OMIM两个数据库检索与AD相关的靶点。对交泰丸和AD的共同靶点进行基因本体(Gene ontology,GO)功能富集分析和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。利用Cytoscape 3.2.0软件构建交泰丸-成分-靶点网络,PPI网络和靶点-功能网络,并对这些网络进行拓扑学分析。同时采用APP_(swe)/PS1_(dE9)双转基因小鼠构建AD动物模型,以中药复方交泰丸灌胃处理后,采用Morris水迷宫测试小鼠的空间记忆能力,用检测试剂盒检测小鼠脑组织内氧化应激水平,并通过Western Blot法检测相关蛋白,以初步验证网络药理学预测结果。结果:检索黄连和肉桂2味中药,筛选得到黄连有效化学成分14个,肉桂有效化学成分7个,共得到21个有效化学成分。对这些化学成分进行靶点预测,共得到295个交泰丸靶点。对AD相关靶点进行检索,共得到912个与AD相关的靶点。GO功能富集分析表明交泰丸可能对学习记忆、神经功能、细胞死亡、酶活性、炎症、免疫功能、氧化应激、线粒体功能、细胞生长和代谢有影响。KEGG通路富集分析表明交泰丸治疗AD涉及HIF-1信号通路、TNF信号通路、FoxO信号通路、PI3K-AKT信号通路、凋亡信号通路、NF-κB信号通路和MAPK信号通路,等。动物实验验证结果表明,交泰丸4.2 g/kg组的小鼠的空间记忆障碍能力得到改善,可降低APP_(swe)/PS1_(dE9)双转基因小鼠脑组织内氧化应激水平,可激活PI3K/AKT信号通路(P<0.05或P<0.01)。结论:本研究揭示了交泰丸可以通过多成分,多靶点和多途径发挥神经保护作用以治疗阿尔茨海默病,为今后交泰丸的深入研究提供了良好的科学依据。
Objective:The mechanism of Jiaotaiwan in the treatment of Alzheimer’s disease(AD)was investigated through the network pharmacologys,and it was validated by animal experiments.Methods:In this study,the effective chemical constituents of Coptis chinensis and cinnamon in Jiaotaiwan were retrieved by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).After the retrieval of related literature on oral bioavailability(OB)and drug-likeness(DL),the effective chemical constituents were obtained and the target was predicted.AD related targets were retrieved from GeneCards and OMIM databases.The common targets of Jiaotaiwan and AD were analyzed by Gene ontology(GO)functional enrichment and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment.Jiaotaiwan-component-target network,PPI network and target-function network were constructed by Cytoscape 3.2.0 software,and the topological analysis was performed.At the same time,the AD animal model was established by APP_(swe)/PS1_(dE9)transgenic mice.After the intragastric administration of Jiaotaiwan,the Morris water maze was used to test the spatial memory ability of mice,the levels of oxidative stress in brain tissues of mice were detected by the kit,and the related proteins were detected by Western Blot method to preliminarily verify the prediction results of network pharmacology.Results:14 effective chemical constituents of Coptis chinensis and 7 effective chemical constituents of cinnamon were obtained.A total of 21 effective chemical constituents were obtained.A total of 295 targets of Jiaotaiwan were obtained by the target prediction.A total of 912 AD-related targets were obtained.GO functional enrichment analysis showed that Jiaotaiwan had an effect on the learning and memory,the nerve function,the cell death,the enzyme activity,the inflammation,the immune,the oxidative stress,the mitochondrial function,the cell growth and metabolism.Enrichment analysis of KEGG pathway showed that the treatment of AD with Jiaotaiwan involved HIF-1 signaling pathway,TNF signaling pathway,FoxO signaling pathway,PI3 K-Akt signaling pathway,apoptosis signaling pathway,NF-kappa B signaling pathway and MAPK signaling pathway.The animal experiments results showed that the spatial memory impairment ability of mice treated with 4.2 g/kg Jiaotaiwan was improved.4.2 g/kg Jiaotaiwan also reduced the level of oxidative stress in the brain tissue of APP_(swe)/PS1_(dE9)transgenic mice,and activated the PI3 K/Akt signaling pathway.Conclusion:This study reveals that Jiaotaiwan can exert the neuroprotective effects in the treatment of Alzheimer’s disease through multiple-components,multiple-targets and multiple-pathways,which provides a good scientific basis for further research on Jiaotaiwan.
作者
高欣
蔡伟武
魏素芬
罗艺
李伟荣
黄水清
张彦红
王奇
王宏
Gao Xin;Cai Weiwu;Wei Sufen;Luo Yi;Li Weirong;Huang Shuiqing;Zhangyanhong;Wang Qi;Wang Hong(Science and Technology Innovation Center of Guangzhou University of Chinese Medicine,Guangzhou 510405;Laboratory Animal Center of Guangzhou University of Chinese Medicine,Guangzhou 510405;Institute of Clinical Pharmacology,Guangzhou University of Chinese Medicine,Guangzhou 510405;Department of Traditional Chinese Medicine,Guangzhou First People’s Hospital,School of Medicine,South China University of Technology,Guangzhou 510405)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第1期7-13,共7页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金项目(编号:81973918)
广州市中医药防治脑病重点实验室广州市科技局项目(编号:201805010005,201803010047)
广东省自然科学基金项目(编号:2016A030313435)
关键词
交泰丸
阿尔茨海默病
网络药理学
作用机制
实验验证
Jiaotaiwan
Alzheimer’s disease
Network Pharmacology
Mechanism of action
experimental verification
作者简介
通信作者:王宏,研究员,研究方向:中医脑病实验研究,E-mail:zyjw@gzucm.edu.cn;高欣,实验师,学士,研究方向:中医脑病实验研究,E-mail:cocoko@gzucm.edu.cn。
