基于网络药理学和实验验证探究栀子豉汤治疗抑郁症的配伍机制  

A study on the compatibility mechanism based on network pharmacology and experimental validation of Zhi-zi-chi Decoction in the treatment of depression

作　　者：叶莹 何镜 刘筱婧 高玉烨 周婷婷 高晓霞 YE Ying;HE Jing;LIU Xiaojing;GAO Yuye;ZHOU Tingting;GAO Xiaoxia(School of Pharmacy,Guangdong Pharmaceutical University,Guangzhou 510006,China;Department of Pharmacy,People’s Liberation Army Navy Military Medical University,Shanghai 200433,China)

机构地区：[1]广东药科大学药学院,广东广州510006 [2]中国人民解放军海军军医大学药学系,上海200433

出　　处：《沈阳药科大学学报》2024年第5期557-570,共14页Journal of Shenyang Pharmaceutical University

基　　金：国家自然科学基金资助项目(81973457);上海市科委“科技创新行动计划”(21S21902400);上海市卫生健康委卫生健康学科带头人培养计划(2022XD037)。

摘　　要：目的基于网络药理学和体内外实验验证探究栀子豉汤治疗抑郁症的配伍机制。方法采用UPLC-QE-MS技术分析栀子豉汤及其组方药材栀子、淡豆豉中化学成分,结合网络药理学预测栀子、淡豆豉、栀子豉汤治疗抑郁症的相关靶点和信号通路;通过体内外实验对预测结果进行验证:采用抗生素鸡尾酒疗法建立伪无菌大鼠模型,寻找栀子、淡豆豉和栀子豉汤在体内肠道菌群影响下的入血成分及其代谢物的差异;利用谷氨酸诱导的PC12细胞模型评价栀子、淡豆豉和栀子豉汤含药血清以及入血成分基于神经保护的抗抑郁作用。结果分别在栀子、淡豆豉、栀子豉汤中鉴定出37、7和43种化学成分,确定了栀子、淡豆豉、栀子豉汤治疗抑郁症的187、146和196个潜在靶点,从而进一步发现栀子和栀子豉汤治疗抑郁症的9个共同核心靶点;富集分析结果显示,栀子、栀子豉汤主要通过神经活性配体-受体通路、MAPK信号通路和IL-17信号通路等多条信号通路发挥抗抑郁作用。对栀子、淡豆豉、栀子豉汤体内入血成分进行分析,栀子豉汤给药正常大鼠后可鉴定出18种入血成分及其代谢物,而栀子、淡豆豉给药正常大鼠,栀子豉汤给药伪无菌大鼠分别鉴定出9、3和12种入血成分及其代谢物,说明栀子在没有淡豆豉配伍或无体内肠道菌群调控下,其体内代谢会发生改变;体外细胞实验表明各组含药血清及部分入血成分具有神经保护作用,其中栀子豉汤含药血清以及栀子新苷的神经保护作用最显著,各含药血清均可下调谷氨酸诱导的PC12细胞中促炎细胞因子IL-1β、TNF-α和IL-6水平升高。结论淡豆豉具有一定的神经保护作用,与栀子配伍后能够调控体内肠道菌群,影响栀子的体内代谢,栀子豉汤的抗抑郁作用可能与其抑制炎症反应有关。Objective Based on network pharmacology and in vivo and in vitro experimental validation to explore the action mechanism of Zhi-zi-chi Decoction(ZZCD)in the treatment of depression.Methods The UHPLC-QE-MS technique was used to analyze the components in Gardeniae Fructus(Zhizi in Chinese,ZZ in brief),Sojae Semen Praeparatum(Dandouchi in Chinese,DDC in brief)and ZZCD,combined with bioinformatics to predict the relevant targets and signalling pathways of ZZ,DDC and ZZCD for depression treatment.The predictions were validated by in vivo and in vitro experiments.The pseudo germ-free rats were established using antibiotic cocktail(Abx)pretreatment to find the differences in blood entry components and their metabolites under the influence of intestinal flora in vivo for ZZ,DDC and ZZCD.The antidepressant effects of ZZ,DDC and ZZCDcontaining serum based on neuroprotection were also evaluated using the glutamate-induced PC12 cell model.Results In ZZ,DDC and ZZCD,37,7 and 43 components were identified respectively,187,146 and 196 potential targets were identified for the treatment of depression,respectively.And 9 common core targets were further screened.The results of enrichment analysis showed that ZZ and ZZCD exerted antidepressant effects through multiple signaling pathways such as neuroactive ligandreceptor pathway,MAPK signaling pathway and IL-17 signaling pathway.In the analysis of ZZ and ZZCD blood components,18 blood components and their metabolites were identified in ZZCD-administered normal rats,whereas 9,3 and 12 blood components and their metabolites were identified in ZZ,DDC-administered normal rats and ZZCD-administered pseudo germ-free rats.The above results suggested that ZZ in vivo metabolism was altered in the absence of DDC compatibility or in the absence of in vivo intestinal flora modulation.Celluar experiments showed that both ZZ and ZZCD had neuroprotective effects,with the most significant neuroprotective effect of ZZCD-containing serum.The levels of pro-inflammatory cytokines IL-1β,TNF-αand IL-6

关 键 词：栀子豉汤 栀子 淡豆豉 抑郁症 肠道菌群 

分 类 号：R96[医药卫生—药理学] R28[医药卫生—药学]