基于网络药理学分析加味归脾汤治疗阴火失眠伴焦虑的机制  

Analysis on Mechanisms of Modified Guipitang in Treatment of Yin-Fire Insomnia with Anxiety Based on Network Pharmacology

作　　者：刘飞祥 林子璇 张怀亮[1] 杨克勤[1] 张道培 徐进[1] 范晓飞[1] 王永涛[1] 李丹[1] 赵玉男 LIU Fei-xiang;LIN Zi-xuan;ZHANG Huai-liang;YANG Ke-qin;ZHANG Dao-pei;XU-Jin;FAN Xiao-fei;WANG Yong-tao;LI Dan;ZHAO Yu-nan(The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;School of Medicine and Life Sciences,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]河南中医药大学第一附属医院,郑州450000 [2]南京中医药大学医学与生命科学学院,南京210023

出　　处：《中国实验方剂学杂志》2020年第20期161-168,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：全国名老中医药专家传承工作室建设项目(国中医药人教函[2018]134号);吴阶平医学基金会临床科研专项(320.6750.18326)。

摘　　要：目的:借助网络药理学分析技术,探讨加味归脾汤治疗阴火失眠伴焦虑的作用机制。方法:采用中药系统药理学数据库及分析平台(TCMSP),筛选加味归脾汤的主要化合物成分和药物靶基因;通过GeneCards和人类孟德尔遗传综合数据库(OMIM)建立失眠和焦虑的靶基因集;通过STRING 11.0软件分析交集基因之间的相互作用;采用Cytoscape3.6.1软件分析和马修斯相关系数(MCC)算法筛选核心基因。基于网络分析结果,将48只SD雌性大鼠随机分配为空白组,模型组,右佐匹克隆片组(0.2 mg·kg^-1·d^-1),加味归脾汤低、中、高剂量组(0.31,1.25,5 g·kg^-1·d^-1)。采用氯苯丙氨酸(PCPA)腹腔注射方法,建立失眠伴焦虑动物模型,并给予相应药物治疗7 d;然后进行自主活动实验观察大鼠活动的次数、时间和路程;实时荧光定量聚合酶链式反应检测大鼠海马组织中RAC-α丝氨酸/苏氨酸蛋白激酶(Akt1),丝裂原活化蛋白激酶3(MAPK3),丝裂原活化蛋白激酶8(MAPK8)和白细胞介素-6(IL-6)mRNA表达。结果:从TCMSP数据库中共筛选出228个化合物活性成分;经与GeneCards和OMIM综合数据库比对,共获取181个疾病与药物的交集基因;STRING软件和MCC算法共获得MAPK3,MAPK8,Akt1,IL-6等9个关键基因。动物实验表明,与模型组比较,加味归脾汤高、中剂量组的活动次数、时间和路程显著减少(P<0.01),海马组织中MAPK3,MAPK8,Akt1,IL-6 mRNA表达显著降低(P<0.01),低剂量组差异无统计学意义。结论:加味归脾汤治疗阴火失眠伴焦虑作用机制,可能与对MAPK3,MAPK8,Akt1,IL-6基因的调节有关。Objective: To explore the mechanism of modified Guipitang in the treatment of Yin-Fire insomnia with anxiety with the help of network pharmacological analysis technology. Method: Traditional Chinese Medicine Systems Pharmacology(TCMSP)was used to screen the main components and target genes of modified Guipitang. GeneCards and Online Mendelian Inheritance in Man(OMIM)were used to establish the target gene sets of insomnia and anxiety. STRING 11.0 software was used to analyze the interaction between the overlapping genes, and Cytoscape3.6.1 software analysis and Matthews correlation coefficient(MCC)algorithm were used to screen the core genes. Based on the results of network analysis,48 SD female rats were randomly divided into blank control group,model group,eszopiclone tablets group(0.2 mg·kg^-1·d^-1),modified Guipitang low,medium,and high-dose groups(0.31,1.25,5 g·kg^-1·d^-1). The model of insomnia with anxiety was established by intraperitoneal injection of Para-chlorophenylalanine(PCPA)and these rats were treated with corresponding drugs for 7 days. Then the frequency,time and distance of the activities were observed in the experiment of autonomic activity. Real-time quantitative polymerase chain reaction(PCR)was used to detect the mRNA expressions of proactivated protein kinase 8(MAPK8),RAC-alpha serine/threonine protein kinase(Akt1),mitogen-activated protein kinase 3(MAPK3)and interleukin-6(IL-6)in rat hippocampus. Result:A total of 228 active compounds were screened from TCMSP database and 181 intersecting genes of diseases and drugs were obtained by comparing with GeneCards and OMIM comprehensive database. 9 core genes,including MAPK3, MAPK8, Akt1 and IL-6 were identified by STRING software and MCC algorithm. Animal experiments showed that the number of activity times,time and distance of modified Guipitang in high and medium dose groups were significantly lower than those in the model group. The high and middle dose groups of modified Guipitang could significantly inhibit the mRNA expression of MAPK3,

关 键 词：网络药理学 加味归脾汤 失眠 焦虑 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]