摘要
目的:基于生物信息学方法探索橙皮苷治疗抑郁症的作用机制,并通过动物实验进行初步验证。方法:通过DisGeNET、Drugbank、GeneCards数据库及GSE76826数据集筛选抑郁症的靶点。借助TCMIP、TCMSP、BATMAN-TCM及Swiss Target Prediction数据库检索橙皮苷的靶点。将抑郁症与橙皮苷的靶点取交集,并绘制韦恩图。利用STRING数据库绘制蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,并筛选关键靶点。采用R语言对关键靶点进行京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路分析,并筛选关键通路。通过AutoDock 1.5.6软件对橙皮苷与关键靶点进行分子对接分析。采用R语言对关键靶点进行ROC分析,并获取曲线下面积(area under concentration-time curve,AUC)。将24只SD大鼠随机分为正常组、模型组及橙皮苷组(20 mg·kg^(-1)),每组8只。采用慢性束缚法建立抑郁症模型,持续21 d。采用旷场实验、糖水偏好实验及高架十字迷宫实验评价大鼠的抑郁样行为。采用超高效液相色谱(ultra performance liquid chromatography,UPLC)法检测大鼠前额叶皮质中去甲肾上腺素(norepinephrine,NE)、肾上腺素(epinephrine,EP)、左旋多巴(levodopa,L-DOPA)、多巴胺(dopamine,DA)、5-羟色胺(5-hydroxytryptamine,5-HT)、高香草酸(homovanillic acid,HVA)含量。结果:抑郁症靶点3850个,橙皮苷靶点149个,两者的交集靶点83个。12个关键靶点涉及的KEGG通路包括PI3K/AKT信号通路、MAPK信号通路及TNF信号通路等。其中,PI3K/AKT信号通路可能是橙皮苷治疗抑郁症的关键通路。5个关键靶点与橙皮苷的分子对接结合能均小于-7.0 kcal·mol^(-1)。ROC分析显示,TP53与VEGFA的诊断价值较高(AUC>0.900)。模型组大鼠的中央区停留时间、穿格次数、直立次数、运动总距离较正常组减少(P<0.01),经橙皮苷治疗后增加(P<0.05)。模型组大鼠的糖水偏好率、开臂进入次数百分比、开臂进入时间百分比较正常组降低,经橙皮苷治疗后升高(P<0.05)。模型组大鼠前额叶皮质中NE、EP、L-DOPA、DA、5-HT、HVA含量较正常组减少,经橙皮苷组治疗后增加(P<0.01)。结论:橙皮苷可能通过调控PI3K/AKT信号通路,提高前额叶皮质单胺类神经递质含量,从而发挥抗抑郁作用。
Objective:To explore the mechanism of hesperidin in the treatment of depression based on bioinformatics methods,and to conduct preliminary verification through animal experiments.Methods:DisGeNET,Drugbank,GeneCards databases and GSE76826 datasets were used to screen the targets of depression.Hesperidin targets were retrieved using TCMIP,TCMSP,BATMAN-TCM and Swiss Target Prediction databases.The targets of depression and hesperidin were intersected,and a Wayne diagram was plotted.The STRING database was used to map the protein-protein interaction(PPI)network and screen key targets.The Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis of key targets was performed using R language,and the key pathways were screened.Molecular docking analysis of hesperidin to key targets was performed by Auto Dock 1.5.6 software.ROC analysis of key targets was performed in R language,and the area under concentration-time curve(AUC)was obtained.Twenty-four SD rats were randomly divided into blank group,model group and hesperidin group(20 mg·kg^(-1)),with 8 rats in each group.The chronic restraint method was used to establish a depression model for 21 days.The open field experiment,sugar water preference test and elevated cross maze test were used to evaluate the depressive behavior of rats.Ultra performance liquid chromatography(UPLC)was used to detect norepinephrine(NE),epinephrine(EP),levodopa(L-DOPA),dopamine(DA),5-Serotonin(5-hydroxytryptamine)(5-HT)and homovanillic acid(HVA)content.Results:There were 3850 targets for depression,149 targets for hesperidin,and 83 targets at the intersection of the two.The KEGG pathways involved in the 12 key targets include PI3K/AKT signaling pathway,MAPK signaling pathway and TNF signaling pathway.Among them,the PI3K/AKT signaling pathway may be a key pathway for hesperidin in the treatment of depression.The molecular docking binding energy of the five key targets with hesperidin was less than-7.0 kcal·mol^(-1).ROC analysis showed that TP53 and VEGFA had a higher diagnostic value(AUC>0.900).Compared with the normal group,the central dwell time,the number of penetrations,the number of erections,and the total exercise distance of the model group decreased(P<0.01)and increased after hesperidin treatment(P<0.05).The sugar water preference rate,the percentage of open-arm entry times,and the percentage of open-arm entry time in the model group were lower than those in the normal group and increased after hesperidin treatment(P<0.05).The contents of NE,EP,L-DOPAC,DA,5-HT and HVA in the prefrontal cortex of the model group decreased compared with the normal group and increased after treatment with hesperidin group(P<0.01).Conclusion:Hesperidin may exert antidepressant effect by regulating the PI3K/AKT signaling pathway and increasing the content of monoamine neurotransmitters in the prefrontal cortex.
作者
吴永叶
杜志欣
杨丽萍
侯俊林
余晨阳
王昱龙
王钰洁
刘玉娟
WU Yongye;DU Zhixin;YANG Liping;HOU Junlin;YU Chenyang;WANG Yulong;WANG Yujie;LIU Yujuan(Traditional Chinese Medicine School(Zhongjing School),Henan University of Chinese Medicine,Zhengzhou Henan China 450046)
出处
《中医学报》
2025年第5期1091-1100,共10页
Acta Chinese Medicine
基金
国家自然科学基金项目(81973596)
河南省大学生创新创业训练计划项目(S202210471038)。
