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目的 基于UPLC-MS/MS和网络药理学探讨左归丸抗阿尔茨海默病(AD)药效物质及作用机制。方法 采用UPLC-MS/MS法鉴定左归丸的化学成分；采用网络药理学方法分析左归丸的有效成分、作用靶点及信号通路；采用分子对接技术分析左归丸活性成分与作用靶点的结合情况；噻唑蓝比色法(MTT)测定左归丸对HT22细胞增殖的影响；酶联免疫吸附法(ELISA)检测AD细胞模型的Aβ1-42表达水平；蛋白免疫印迹法检测p-PI3K和p-AKT蛋白表达水平。结果 UPLC-MS/MS法共鉴定出501种化合物，通过HERB和SwissADME数据库鉴定出22种有效成分。通过与AD相关的靶点取交集，确定了123个交集靶点。从蛋白质互作网络(PPI)中获得8个核心靶点。GO富集分析结果表明，这些靶点主要与外源刺激、基因表达的正调控、对缺氧的反应有关。KEGG通路分析表明，这些靶点与癌症通路、神经变性-多发性疾病通路、阿尔茨海默病通路和PI3K/AKT信号通路相关。分子对接结果表明，左归丸有效成分与磷脂酰肌醇-3激酶催化亚基α(PIK3CA)、表皮生长因子受体(EGFR)、基质金属蛋白酶9(MMP9)、雷帕霉素靶蛋白(mTOR)和肿瘤坏死因子(TNF)、丝裂原激活蛋白激酶激酶1(MAP2K1)、RAF激酶蛋白(BRAF)具有良好的结合亲和力。体外实验证明，中、高浓度左归丸可显著提升细胞活力，降低HT22细胞Aβ1-42水平(P<0.05)，提高p-PI3K和p-AKT蛋白的表达水平(P<0.05)。结论 左归丸含有22种有效成分，其可能通过调节PI3K/AKT通路，减少Aβ1-42的产生，进而改善AD。
Abstract：Objective To determine the pharmacodynamic substances and mechanism of Zuogui pills (ZGP) in Alzheimer’s disease (AD) based on UPLC-MS/MS and network pharmacology.Methods UPLC-MS/MS was used to identify the chemical compounds in ZGP.Network pharmacology was used to analyze the active ingredients,targets and signal pathways of ZGP.Molecular docking was used to analyze the binding of ZGP’s active ingredients and AD related targets.MTT was used to determine the effect of ZGP on the proliferation of HT22 cells,ELISA to detect the expression level of Aβ1-42 in AD cell models and Western blot to detect the expression levels of p-PI3K and p-AKT proteins.Results Totally 501 compounds were identified with UPLC-MS/MS,22 active ingredients were identified via HERB and SwissADME databases.Totally 123 common targets were identified by intersecting with AD-related targets,and 8 core targets were obtained from the PPI network.The GO enrichment analysis showed that these targets were mainly associated with exogenous stimuli,positive regulation of gene expression,and response to hypoxia.KEGG pathway analysis showed that these targets were also associated with the cancer pathway,neurodegenerative-multiple disease pathway,Alzheimer’s disease pathway,and PI3K/AKT signaling pathway.Molecular docking revealed that active ingredients of ZGP had good binding affinity to PIK3CA,EGFR,MMP9,MTOR,TNF,MAP2K1 and BRAF.In vitro experiments demonstrated that medium and high concentrations of ZGP significantly enhanced the cell viability,reduced Aβ1-42 clustering in HT22 cells (P<0.05),and elevated the expression levels of p-PI3K and p-AKT proteins (P<0.05).Conclusion ZGP contains 22 active ingredients,which may reduce the production of Aβ1-42 by regulating the PI3K/AKT pathway,and improve AD symptoms.
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基本信息:
DOI：
中图分类号:R285.5
引用信息:
[1]王燕,唐亮,刘瑶等.基于UPLC-MS/MS、网络药理学、分子对接和体外实验探究左归丸治疗阿尔茨海默病的机制[J].中南药学,2025,23(08):2157-2164.
基金信息:
湖南省卫健委科研计划课题重点指导项目(No.C202303078708); 湖南省教育厅科学研究项目重点项目(No.22A0025);湖南省教育厅科研项目(优秀青年)(No.23B0875); 长沙市自然科学基金(No.kq2208175)