UHPLC-TOF-MS结合网络药理学与实验验证探讨左金丸治疗抑郁症作用机制  

Anti-depression mechanism of Zuojin Pills:based on UHPLC-TOF-MS, network pharmacology, and experimental verification

作　　者：戴国梁[1] 杭华茜 陈佩瑶 洪盛威 许美娟[1] 马程遥 黄茜 叶峪 于美双 居文政[1] DAI Guo-liang;HANG Hua-xi;CHEN Pei-yao;HONG Sheng-wei;XU Mei-juan;MA Cheng-yao;HUANG Qian;YE Yu;YU Mei-shuang;JU Wen-zheng(Department of Clinical Pharmacology,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China)

机构地区：[1]南京中医药大学附属医院临床药理科,江苏南京210029

出　　处：《中国中药杂志》2023年第1期183-192,共10页China Journal of Chinese Materia Medica

基　　金：江苏省中医药科技发展计划面上项目(MS2021011);江苏省基础研究计划面上项目(BK20211394,BK20221425);江苏省中医药领军人才项目(SLJ0208)。

摘　　要：基于入血成分、网络药理学与实验验证研究策略,探讨左金丸治疗抑郁症的作用机制。采用UHPLC-TOF-MS对左金丸含药血清进行定性分析。采用PharmMapper、GeneCards等数据库获取入血成分及疾病相关靶点,取交集后构建蛋白互作(PPI)网络,筛选核心靶点并进行GO功能和KEGG通路富集分析。采用Cytoscape 3.7.2软件构建“化合物-靶点-通路”网络,预测左金丸抗抑郁的作用靶点与信号通路,并构建CUMS抑郁模型小鼠对关键靶点进行验证。结果显示,共鉴定左金丸入血成分21个,主要为生物碱类化合物。化合物-疾病共有靶点155个,PPI网络分析筛选出核心靶点67个。KEGG富集与PPI网络分析发现,左金丸可能通过AMPK/SIRT1、NLRP3、insulin等靶点与通路发挥抗抑郁作用。动物实验结果表明,左金丸可显著改善CUMS抑郁模型小鼠的抑郁样行为,显著降低海马区和血清中IL-1β、IL-6及TNF-α水平,激活AMPK/SIRT1信号,并降低NLRP3的蛋白表达水平。综上可得,左金丸可能主要通过激活小鼠海马区AMPK/SIRT1信号通路,抑制NLRP3激活及神经炎症,从而发挥抗抑郁作用。This study aims to explore the anti-depression mechanism of Zuojin Pills based on the plasma constituents, network pharmacology, and experimental verification. UHPLC-TOF-MS was used for qualitative analysis of Zuojin Pills-containing serum. Targets of the plasma constituents and the disease were retrieved from PharmMapper and GeneCards. Then the protein-protein interaction(PPI) network was constructed and core targets were screened for GO term enrichment and KEGG pathway enrichment. Cytoscape 3.7.2 was employed construct the "compound-target-pathway" network and the targets and signaling pathways of Zuojin Pills against depression were predicted. CUMS-induced depression mouse model was established to verify the key targets. The results showed that a total of 21 constituents migrating to blood of Zuojin Pills were identified, which were mainly alkaloids. A total of 155 common targets of the constituents and the disease and 67 core targets were screened out. KEGG enrichment and PPI network analysis showed that Zuojin Pills may play a role in the treatment of depression through AMPK/SIRT1, NLRP3, insulin and other targets and pathways. Furthermore, the results of animal experiments showed that Zuojin Pills could significantly improve the depression behaviors of depression, reduce the levels of IL-1β, IL-6 and TNF-α in hippocampus and serum, activate AMPK/SIRT1 signaling, and reduce the protein expression of NLRP3. In conclusion, Zuojin Pills may play a role in the treatment of depression by activating AMPK/SIRT1 signaling pathway, and inhibiting NLRP3 activation and neuroinflammation in the hippocampus of mice.

关 键 词：左金丸 抑郁症 网络药理学 UHPLC-TOF-MS 小鼠 

分 类 号：R285[医药卫生—中药学]