| 马澜,时晶,李傅尧,等.中药复方金思维肠吸收液治疗阿尔茨海默病的药效物质基础及作用机制研究[J].浙江中医药大学学报,2024,48(12):1467-1481. |
| 中药复方金思维肠吸收液治疗阿尔茨海默病的药效物质基础及作用机制研究 |
| Research on the Pharmacodynamic Material Basis and Mechanism of Action of the Chinese Medicine Compound GAPT Intestinal Absorption Solution in Treating Alzheimer’s Disease |
| DOI：10.16466/j.issn1005-5509.2024.12.001 |
| 中文关键词: 阿尔茨海默病 金思维 PI3K/AKT/GSK-3β信号通路 人神经母细胞瘤细胞 HPLC-MS技术 肠吸收液 网络药理学 痴呆 |
| 英文关键词: Alzheimer’s disease GAPT PI3K/AKT/GSK-3β signaling pathway human neuroblastoma cells HPLC-MS intestinal absorption solution network pharmacology dementia |
| 基金项目:国家自然科学基金项目(82074362)；北京中医药大学“解码中医”协同攻关项目(BZY-JMZY-2022-002) |
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| 中文摘要: |
| [目的] 测定中药复方金思维肠吸收液的有效成分并进行网络药理学分析，探究其治疗阿尔茨海默病(Alzheimer’s disease，AD)的作用机制并进行实验验证。[方法] 采用高效液相质谱色谱联合(high performance liquid chromatography-tandem mass spectrometry，HPLC-MS)法测定金思维肠吸收液和金思维原液的共有成分，使用中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform，TCMSP)对共有成分进行筛选，使用SWISS数据库查询药物作用靶点；使用DrugBank、OMIM、GeneCards、DisGeNET、TTD数据库查询AD靶点。对药物靶点与疾病靶点进行韦恩分析，得到金思维治疗AD的关键靶点。使用STRING数据库进行蛋白互作(protein-protein interaction，PPI)分析，使用Cytoscape 3.7.2软件绘制PPI网络图。将关键靶点导入Metascape平台，进行基因本体(gene ontology，GO)分析和京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes，KEGG)通路分析，使用Cytoscape 3.7.2软件将相关网络关系可视化。最后依据网络药理学分析结果，选择磷脂酰肌醇3-激酶/蛋白激酶B (phosphoinositide 3-kinase/protein kinase B，PI3K/AKT)信号通路进行细胞实验验证。[结果] 共筛选出符合条件的活性成分73个，其中延胡索甲素、甜橙黄酮、黄芩黄酮Ⅱ、甘草素、黄藤素为关联靶点数量排名前5的活性成分，即金思维肠吸收液治疗AD的关键成分。PPI网络分析提示，AKT1等为治疗AD的核心靶点。KEGG分析提示，金思维肠吸收液治疗AD可能涉及PI3K/AKT信号通路、钙信号通路、环磷酸腺苷(cyclic adenosine monophosphate，cAMP)信号通路等。细胞实验结果表明，模型组磷酸化AKT(phosphorylated-AKT，p-AKT)蛋白表达较正常组显著降低(P<0.01)；模型组磷酸化糖原合酶激酶-3β/糖原合酶激酶-3β(phosphorylated-glycogen synthase kinse-3β/glycogen synthase kinse-3β，p-GSK-3β/GSK-3β)比值较正常组显著降低(P<0.05)，金思维肠吸收液中、高剂量组较模型组显著升高(P<0.0001，P<0.001)，通路抑制剂组与模型组差异无统计学意义(P>0.05)。[结论] 金思维肠吸收液通过多途径、多靶点治疗AD，其中对PI3K/AKT信号通路及其下游靶点的调节是其发挥作用的关键。 |
| 英文摘要: |
| [Objective] To determine the main components of Chinese medicine compound GAPT intestinal absorption solution and conduct network pharmacology analysis， and explore its mechanism of action in treating Alzheimer’s disease(AD) and conduct experimental verification. [Methods] The common components of GAPT intestinal absorption solution and GAPT stock solution were determined by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS)， the common components were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) database， the drug targets were queried by SWISS database， and the AD targets were queried by DrugBank， OMIM， GeneCards， DisGeNET and TTD databases. Venn analysis of drug targets and disease targets yielded the key targets of GAPT for the treatment of AD. Protein-protein interaction(PPI) analysis was performed using the STRING database and PPI network was plotted using Cytoscape 3.7.2 software. Key targets were imported into the Metascape platform for gene ontology(GO) analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis. Correlation network relationships were visualized using Cytoscape 3.7.2 software. Finally， according to the results of network pharmacology analysis， the phosphoinositide 3-kinase/protein kinase B(PI3K/AKT) signaling pathway was selected for cell experimental verification. [Results] A total of 73 eligible active ingredients were screened， among which Corydaline， Sinensetin， Skullcapflavone Ⅱ， Liquiritigenin， Palmatine were key components in the treatment of AD with GAPT intestinal absorption solution. PPI network analysis suggested that AKT1 was the core targets for preventing and treating AD. KEGG analysied suggests that the prevention and treatment of AD with GAPT intestinal absorption solution may involve PI3K/AKT signaling pathway， calcium signaling pathway， cyclic adenosine monophosphate(cAMP) signaling pathway， etc. The results of cell experiments showed that the proportion of phosphorylated-AKT(p-AKT) protein was significantly reduced in model group compared to normal group(P<0.01); the expression level of phosphorylated-glycogen synthase kinse-3β/glycogen synthase kinse-3β(p-GSK-3β/GSK-3β) protein in model group was significantly reduced compared to normal group(P<0.05)， while the medium and high-dose groups of GAPT intestinal absorption solution were significantly increased compared to model group(P<0.0001， P<0.001). There was no significant difference between pathway inhibitor group and model group(P>0.05). [Conclusion] GAPT intestinal absorption solution treats AD through multiple pathways and multiple targets， and the regulation of PI3K-AKT signaling pathway and its downstream targets is the key to its role. |
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