基于网络药理学和实验验证探讨丹星止颤方治疗帕金森病作用机制  

Discussion on the mechanism of Danxing Zhichan Prescription in the treatment of Parkinson's disease based on network pharmacology and experimental verification

作　　者：胡周圆 杨一帆 彭涛 胡楠 云烨栋 阴俊 闫咏梅[1,2] 李涛 贾妮[1] Hu Zhouyuan;Yang Yifan;Peng Tao;Hu Nan;Yun Yedong;Yin Jun;Yan Yongmei;Li Tao;Jia Ni(The First Clinical Medical College of Shaanxi University of Chinese Medicine,Xianyang 712046,China;Department of Encephalopathy,Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]陕西中医药大学第一临床医学院,咸阳712046 [2]陕西中医药大学附属医院脑病科,咸阳712000

出　　处：《国际中医中药杂志》2025年第2期205-212,共8页International Journal of Traditional Chinese Medicine

基　　金：国家岐黄学者支持项目(国中医药人教函〔2022〕6号);陕西省教育厅重点实验室项目(19JS022);中医颤病防治方案循证优化与评价研究(303-202190078);陕西省科技厅陕西省重点研发计划重点产业创新链(群)-社会发展领域研究课题(2021ZDLSF04-13)。

摘　　要：目的通过网络药理学与动物实验相结合的方法,探讨丹星止颤方治疗帕金森病的作用机制。方法检索TCMSP、BATMAN数据库获得丹星止颤方活性成分及作用靶点,检索GeneCards、OMIM数据库获得帕金森病作用靶点;通过Venny 2.1.0对药物靶点和帕金森病相关靶点取交集,采用STRING 12.0数据库构建交集靶点PPI网络,采用Cytoscape 3.10.0软件进行拓扑分析,得到丹星止颤方作用于帕金森病的关键靶点;利用微生信平台对关键靶点进行GO功能和KEGG通路富集分析,通过AutoDockTools 1.5.7进行分子对接验证。采用随机数字表法将小鼠分为空白对照组、模型组和丹星止颤方组,每组20只;除空白组外,其余各组小鼠灌胃鱼藤酮制备帕金森病模型;丹星止颤方组灌胃丹星止颤方浓缩液10.5 g/kg,空白组、模型组灌胃生理盐水0.2 ml,连续灌胃21 d;采用Western blot法检测脑组织Akt1、Bcl-2、Bax、α-突触核蛋白(α-Syn)表达。结果获得丹星止颤方靶点和帕金森病靶点的交集靶点359个,核心靶点69个,活性成分185个,主要活性成分包括槲皮素、山柰酚、苯丙氨酸等,关键靶点为AKT1、TP53、TNF、ESR1等。KEGG通路富集分析主要涉及AGE-RAGE、PI3K-Akt及流体剪刀应力与动脉粥样硬化信号通路等。验证实验结果显示,与模型组比较,中药组Bcl-2蛋白表达上调(P<0.01),Bax、Akt1和α-Syn蛋白表达下调(P<0.01)。结论丹星止颤方具有多靶点、多途径治疗帕金森病的作用特点,其机制可能与下调Bax、Akt1、α-Syn蛋白表达,改善脑部供血、调节神经递质平衡、抑制氧化应激反应和促进神经再生等方面有关。Objective To explore the mechanism of Danxing Zhishuang Prescription in the treatment of Parkinson's disease(PD)by combining network pharmacology with animal models.Methods TCMSP,BATMAN database,Genecards,and OMIM databases were retrieved to obtain the active components and action targets of Danxing Zhishuang Prescription.Venny 2.1.0 was used to intersect drug targets and PD related genes,and a protein interaction network of the intersection targets was constructed using the STRING 12.0 platform.Topology analysis was performed using Cytoscape 3.10.0 software to identify the key targets of Danxing Zhishuang Prescription on PD;GO functional and KEGG pathway enrichment analysis was performed on key targets using the WeChat platform,and molecular docking was validated through AutoDockTools 1.5.7.Using a random number table method,mice were divided into a blank control group,a model group,and a Danxing Zhishuang Prescription group,with 20 mice in each group;except for the blank group,all other groups of mice were orally administered fisetin to prepare PD models;Danxing Zhishuang Prescription group was orally administered with concentrated Danxing Zhishuang Prescription at a dosage of 10.5 g/kg,while the blank group and model group were orally administered with 0.2 ml of physiological saline for 21 days;Western blot was used to detect the expressions of Akt1,Bcl-2,Bax,andα-Syn proteins.Results 359 intersection targets,69 core targets,and 185 active components were obtained the treatment of PD with Danxing Zhishuang Prescription.The main active components included quercetin,kaempferol,phenylalanine,etc.,and the key targets were AKT1,TP53,TNF,ESR1,etc.KEGG analysis revealed several key signaling pathways,such as AGE-RAGE,PI3K-Akt,fluid shear stress and atherosclerosis signaling pathways.The validation experiment results showed that compared with the model group,the expression of Bcl-2 protein was up-regulated(P<0.01),and the expressions of Bax,Akt1,andα-Syn proteins were down-regulated in the Danxing Zhishuang Prescrip

关 键 词：帕金森病 网络药理学 丹星止颤方 小鼠 

分 类 号：R285.5[医药卫生—中药学]