基于网络药理学研究四物汤对阿尔茨海默病的作用机制及实验验证

Siwutang in Treating Alzheimer′s Disease:Mechanism Analysis Based on Network Pharmacology and Experimental Verification

目的:基于网络药理学方法,探讨四物汤治疗阿尔茨海默病(AD)的作用机制。方法:通过中药系统药理学数据库(TCMSP)得到四物汤中药物的化学成分,并预测其作用靶点,使用Cytoscape 3.7.2构建成分-靶点图,从GeneCard和Omim数据库得到AD的疾病靶点,构建蛋白质相互作用网络(PPI),进行基因本体(GO)与基因相互作用(KEGG)富集分析;并通过体外细胞实验进行初步验证。结果:网络药理学研究共得到四物汤活性成分19个,潜在靶点46个。构建PPI网络并进行拓扑学分析,前5位核心靶点有蛋白激酶B1(AKT1)、胱天蛋白酶3(CASP3)、JUN原癌基因(JUN)、肿瘤坏死因子(TNF)、前列腺素内过氧化物合酶2(PTGS2)。富集结果显示,其主要与神经元凋亡过程的正调控等生物过程有关,可能作用于细胞凋亡、Toll样受体(TLRs)信号通路等。体外实验显示,四物汤能够显著增加B细胞淋巴瘤-2(bcl-2)的表达。结论:本研究说明了四物汤对AD多靶点、多通路的整体调节特点,总结了应用四物汤的理论依据,初步揭示了可能的分子机制,为后续的临床应用和实验研究以更进一步的理论支持。

Objective:To investigate the mechanism of Siwutang in the treatment of Alzheimer′s disease(AD)based on network pharmacology.Methods:The chemical components of Siwutang were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),and the corresponding targets were predicted.Cytoscape3.7.2 was used to build the component-target network.The targets of AD were obtained from GeneCards and Online Mendelian Inheritance in Man(OMIM).The protein-protein interaction(PPI)network was established,and the Gene Ontology(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were performed.The prediction results were preliminarily verified by in vitro cell experiments.Results:A total of 19 active components and 46 potential targets of Siwutang were predicted.The PPI network showed that the core targets were protein kinase B1(AKT1),caspase-3(CASP3),Jun proto-oncogene(JUN),tumor necrosis factor(TNF),and prostaglandin-endoperoxide synthase 2(PTGS2).The enrichment results showed that the core targets were mainly involved in the biological processes such as the positive regulation of neuronal apoptosis and the cell apoptosis and Toll-like receptor signaling pathways.The cell experiments showed that Siwutang significantly up-regulated the expression of B-cell lymphoma-2(Bcl-2).Conclusion:Siwutang can treat AD by acting on multiple targets and pathways.This study summarizes the theoretical basis of application of Siwutang and preliminarily reveals the possible molecular mechanism,providing theoretical support for subsequent clinical application and experimental studies.