何首乌-石菖蒲药对治疗阿尔茨海默病网络药理学研究及实验验证

Network Pharmacology Research and Experimental Calidation of Polygonimultiflori Radix-Acori Tatarinowii Rhizoma in Treating Alzheimer Disease

目的应用网络药理学方法预测何首乌-石菖蒲药对治疗阿尔茨海默病的作用靶点及相关信号通路,探讨其作用机制。方法通过中药系统药理学数据库与分析平台(TCMSP)检索并筛选何首乌、石菖蒲的潜在活性成分,并查询活性成分作用靶点,采用DrugBank数据库和Pharmmapper服务器预测有效化学成分的作用靶点,采用Cytoscape3.7.1软件构建活性成分-靶点网络;通过DrugBank、DisGeNET、HPO数据库检索阿尔茨海默病相关基因;将活性成分作用靶点和阿尔茨海默病相关基因进行比对,得到交集靶点,利用STRING数据库构建蛋白相互作用网络;使用Cytoscape3.7.1软件根据度值、介数和紧密度筛选关键靶点,应用Metascape数据库分析关键靶点的KEGG信号通路和GO分子功能;通过MTT、ELISA和qPCR实验验证何首乌-石菖蒲对PC12细胞的保护作用。结果共筛选出何首乌-石菖蒲药对有效成分15个,对应靶点280个,与阿尔茨海默病交集靶点共123个,12个关键靶点的前10条KEGG通路和前20个GO分子功能涉及炎症反应和凋亡等。细胞实验表明,何首乌-石菖蒲药对能有效提高PC12细胞存活率,抑制细胞炎症反应,并抑制p38MAPK/NF-κB信号通路激活。结论何首乌-石菖蒲药对的多种有效成分以协同方式与多个靶点、多种途径相互作用,主要通过抑制炎症反应、抗凋亡等对阿尔茨海默病发挥重要治疗作用。

Objective To predict the targets and related signaling pathways of Polygonimultiflori Radix-Acori Tatarinowii Rhizoma in the treatment of Alzheimer disease and explore the mechanism by network pharmacology method.Methods The potential active components of Polygonimultiflori Radix and Acori Tatarinowii Rhizoma were searched and screened through TCMSP databases,and queried the target points of the active ingredients.The targets of the effective chemical constituents of Polygonimultiflori Radix-Acori Tatarinowii Rhizoma was predicted by DrugBank database and Pharmmapper server,and the active components-targets network was construct by Cytoscape 3.7.1.Alzheimer disease related genes were retrieved through DrugBank,DisGeNET,HPO databases.The intersection target was obtained by comparing the target of the active ingredient with the genes related to Alzheimer disease,and the protein interaction network was constructed by the STRING databases.Cytoscape 3.7.1 software was used to screen key targets based on degree value,betweenness and tightness,and Metascape database was used to analyze the KEGG signal pathway and GO molecular function of key targets.The protective effect of Polygonimultiflori Radix-Acori Tatarinowii Rhizoma on PC12 cells was verified by MTT,ELISA and qPCR experiments.Results A total of 15 active components of Polygonimultiflori Radix-Acori Tatarinowii Rhizoma were screened,corresponded to 280 targets,included 123 targets that intersect with Alzheimer disease,the top 10 KEGG signaling pathways and the top 20 GO molecular functions of 12 key target involved inflammation,apoptosis,etc.Cell experiments also proved that Polygonimultiflori Radix-Acori Tatarinowii Rhizoma could effectively improved the cell survival rate of PC12 cells,and inhibited inflammation of PC12 cells and the activation of p38MAPK/NF-κB signaling pathway.Conclusion A variety of active components of Polygonimultiflori Radix-Acori Tatarinowii Rhizoma interact with multiple targets and multiple pathways in a synergistic manner,and play an important role in the treatment of Alzheimer disease mainly by inhibiting inflammation and anti-apoptosis.