基于网络药理学结合大鼠肾上腺嗜铬细胞瘤细胞探讨地黄抗阿尔茨海默病的作用机制

Anti-Alzheimer’s Disease Mechanism of Rehmanniae Radix Based on Network Pharmacology in Combination with Rat Adrenal Pheochromocytoma Cells

目的 基于网络药理学并结合大鼠肾上腺嗜铬细胞瘤细胞(rat pheochromocytoma cell, PC12)分析地黄抗阿尔茨海默病(Alzheimer’sdisease,AD)的主要活性成分及作用机制。方法 查阅文献收集地黄的化学成分,经TCMSP筛选其主要活性成分,并构建“地黄-活性成分-潜在靶点”相互作用网络图和PPI图,通过DAVID数据库进行GO生物学过程富集和KEGG代谢通路富集分析,最后采用MTT法验证活性成分对PC12细胞的保护作用。结果 网络药理学分析预测出地黄主要活性成分15个,作用于AD的相关靶点343个,涉及MAPK信号通路、代谢通路、胰岛素信号通路、PI3K-Akt信号通路等。PC12细胞实验结果表明,通过网络药理学筛选得到的地黄9个活性成分均能不同程度提高PC12细胞的存活率,其中胡萝卜苷、木犀草素、京尼平1-β-D龙胆双糖苷、芹菜素、蔗糖、棉子糖处理的PC12细胞存活率更高(P<0.05)。结论 地黄对AD的作用呈多成分、多靶点、多通路的调节机制,为地黄用于AD的干预和治疗提供参考。

OBJECTIVE To analyze the main active components of Rehmanniae Radix against Alzheimer’s disease(AD)and its mechanism by using network pharmacology combined rat pheochromocytoma cells(PC12). METHODS Literature was reviewed to collect the chemical components of Rehmanniae Radix, the main active components were screened by TCMSP. And constructed the interaction network diagram of “Rehmanniae Radix-active components-potential targets” and protein-protein interaction network diagram. GO biological process enrichment and KEGG metabolic pathway enrichment were analyzed by DAVID database. Finally, MTT method was used to verify the protective effect of active ingredients on PC12 cells. RESULTS Network pharmacological analysis predicted fifteen main active ingredients of Rehmanniae Radix, and 343 targets associated with AD, which involved MAPK signaling pathway, metabolic pathways, insulin signaling pathway, PI3K-Akt signaling pathway,etc. The results of PC12 cells experiments showed that all the nine active ingredients of Rehmanniae Radix obtained by network pharmacological screening could improve the survival rate of PC12 cells to different degrees, among which the survival rate of PC12 cells treated with daucosterol, luteolin, Genipin-1-β-D-gentiobioside, apigenin, sucrose and raffinose was higher(P<0.05).CONCLUSION Rehmanniae Radix has a multi-component, multi-target and multi-pathway regulatory mechanism on AD,which provides reference for the intervention and treatment of AD by Rehmanniae Radix.