基于HPLC指纹图谱及网络药理学探讨黑桑葚防治阿尔兹海默病潜在机制

The mechanism of black mulberry reatment for AD was studied based on HPLC fingerprint and network pharmacology

利用HPLC构建黑桑指纹图谱并分析黑桑的主要活性成分,结合网络药理学和分子对接技术研究黑桑治疗阿尔兹海默病(Alzheimer’s disease,AD)的作用分子机制。HPLC法建立12批黑桑的指纹图谱并对共有峰进行指认识别,进一步采用相似度评价、聚类分析、主成分分析、正交偏最小二乘法判别分析(orthogonal partial least squares discriminant analysis,OPLS-DA)和理想解相似性排序法(technique for order preference by similarity to ideal solution,TOPSIS)对12批黑桑样品进行评价;通过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)和Swiss Target Prediction平台获得黑桑的活性成分与靶点信息。使用Gene Cards数据库和AD专属数据库(Chemogenomics Database For Alzheimer’s Disease)获得AD的疾病靶点信息,得到黑桑治疗AD的潜在作用靶点。通过String(Search Tool for the Retrieval of Interacting Genes/Proteins,String)和Cytoscape3.7.2构建PPI网络图并进行拓扑分析。利用R软件对核心靶点进行GO和KEGG富集分析。通过Auto Dock软件进行模拟分子对接。黑桑HPLC指纹图谱的相似度均>0.96,并指认出3个色谱峰,通过聚类分析将12批样品聚为3类,主成分分析结果与聚类分析结果一致。网络药理学筛选后最终得到12个核心靶点,并对核心靶点进行富集分析得到1480个GO条目和104个KEGG通路。黑桑可能通过核心靶点调节PI3K-Akt信号通路、中性粒细胞外陷等信号通路,进而调控炎症等反应,多靶点-多通路-多途径达到治疗和预防AD的效果。

The fingerprint of black mulberry was characterized using high-performance liquid chromatography(HPLC)to analyze its primary active constituents.Additionally,the molecular mechanism underlying the treatment of Alzheimer’s disease(AD)by black mulberry was investigated through the integration of network pharmacology and molecular docking technology.The fingerprints of 12 batches of black mulberry were established via HPLC,and common peaks were identified.The 12 batches of black mulberry samples were evaluated through similarity evaluation,cluster analysis,principal component analysis,OPLS-DA analysis,and TOPSIS analysis.The active ingredients and target information of black mulberry were obtained from the TCMSP database and the Swiss Target Prediction platform.Disease target information for AD was acquired from the Gene Cards database and AD database to identify potential targets for AD treatment.PPI network diagram was constructed,and topological analysis was performed using the String database and Cytoscape 3.7.2.The core targets were enriched by GO and KEGG using R software.Molecular docking simulations were conducted using Auto Dock software.The HPLC fingerprints of black mulberry samples exhibited a similarity exceeding 0.96,identifying three chromatographic peaks.Cluster analysis categorized the 12 batches of samples into three categories,with principal component analysis results consistent with those of cluster analysis.Twelve core targets were identified after network pharmacological screening,and 1480 GO entries and 104 KEGG pathways were obtained through enrichment analysis of core targets.Black mulberry may regulate the PI3K-Akt signaling pathway,neutrophil exagination and other signaling pathways through core targets,thereby regulating inflammation and other responses,and achieving the effect of treating and preventing AD through multi-target-multi-pathway.