UHPLC-TOF-MS结合网络药理学及实验验证研究交泰丸治疗抑郁症的作用机制  

Mechanism of Jiaotai Pills in treatment of depression by UHPLC-TOF-MS combined with network pharmacology and experimental validation

作　　者：黄茜 戴国梁[1] 杨欣怡 曹阳[1] 居文政[1] HUANG Qian;DAI Guo-liang;YANG Xin-yi;CAO Yang;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》2024年第7期1896-1904,共9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(BK202103940);江苏省中药临床药理医学重点实验室项目(ZDXYS202209)。

摘　　要：利用UHPLC-TOF-MS技术分析交泰丸在正常大鼠体内入血成分,通过网络药理学和动物实验研究交泰丸治疗抑郁症的作用机制。利用UHPLC-TOF-MS技术检测大鼠灌胃交泰丸后的入血成分,结合DisGeNET、SwissTargetPrediction等数据库筛选抑郁症与交泰丸入血成分的交集靶点,构建蛋白-蛋白相互作用(protein-protein interaction,PPI)网络。将关键靶点导入DAVID平台进行基因本体论(Gene Ontology,GO)分析和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路注释,结合成分、靶点和通路,利用Cytoscape 3.9.1软件构建“成分-靶点-通路”网络,预测交泰丸治疗抑郁症的关键靶点及通路。建立大鼠慢性不可预知性温和应激(chronic unpredictable mild stress,CUMS)抑郁模型,进行行为学实验,ELISA法检测大鼠血清炎症因子和脑内神经递质水平,蛋白免疫印迹(Western blot)法检测大鼠海马组织中关键靶点表达。结果鉴定出交泰丸在正常大鼠体内入血成分共17个,10个为生物碱类化合物。入血成分与疾病共有交集靶点124个,根据PPI网络结果筛选出核心靶点52个,其中包括NLRP3和caspase-1等,KEGG富集结果显示主要涉及NOD样受体通路等15条典型通路。动物实验研究结果显示,交泰丸可显著改善CUMS抑郁模型大鼠的抑郁样行为,提高脑内神经递质5-羟色胺(5-hydroxytryptamine,5-HT)、多巴胺(dopamine,DA)及去甲肾上腺素(norepinephrine,NE)的水平,降低血清白细胞介素-1β(interlenkin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)及白细胞介素-6(interlenkin-6,IL-6)水平,下调海马区NLRP3和caspase-1蛋白表达水平,抑制NLRP3介导的NOD样受体信号通路。综上所述,交泰丸可能通过抑制NLRP3炎症小体,抑制NLRP3介导的NOD样受体通路,发挥抗抑郁的作用。This study aims to analyze the constituents of Jiaotai Pills migrating to the blood in normal rats by UHPLC-TOF-MS technique and reveal the underlying mechanism of Jiaotai Pills in the treatment of depression by network pharmacology and animal experiments.UHPLC-TOF-MS technique was used to detect the constituents of Jiaotai Pills in the blood of rats after intragastric administration.The intersection target of the constituents and depression was screened by DisGeNET and SwissTargetPrediction database,and the protein-protein interaction(PPI)network was constructed.Key targets were imported into the DAVID platform for Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway annotation.Combined with constituents,targets,and pathways,the"constituent-target-pathway"network was constructed by Cytoscape 3.9.1 software,through which the key targets and pathways of Jiaotai Pills against depression were predicted.The depression model of chronic unpredictable mild stress(CUMS)was established on rats.After that,behavioral experiments were conducted.The expression of inflammatory factors in serum and the neurotransmitters in the brain were detected by ELISA,and the expression of key targets in the hippocampus was detected by Western blot.The results showed that a total of 17 constituents of Jiaotai Pills were identified in the blood,including 10 alkaloids.There were 124 intersection targets between constituents of Jiaotai Pills and depression disorder.A total of 52 core targets were screened according to PPI results,including NLRP3 and caspase-1,etc.KEGG enrichment analysis mainly involved 15 typical pathways such as NOD-like receptor pathway.The results of animal experiments showed that Jiaotai Pills significantly improved the depression-like behavior of CUMS depressive model on rats,decreased the levels of IL-1β,TNF-αand IL-6 in serum,and increased the expression of neurotransmitters such as 5-hydroxytryptamine(5-HT),dopamine(DA),and norepinephrine(NE)in the brain.Besides,Jiaotai Pills also down-

关 键 词：交泰丸 抑郁症 网络药理学 UHPLC-TOF-MS 炎症 

分 类 号：R285.5[医药卫生—中药学]