摘要
目的 基于病证结合的原则,运用网络药理学的方法分析木丹颗粒治疗气虚血瘀证糖尿病周围神经病变(Diabetic peripheral neuropathy, DPN)的分子机制,并通过分子对接和动物实验加以验证。方法 通过中药系统药理学数据库与分析平台(TCMSP),中医药综合数据库(TCMID)获取木丹颗粒主要活性成分及其靶点,从SymMap数据库中收集气虚血瘀证11个中医症状相应的现代医学症状(MM Symptom)名称,采用人类基因数据库(GeneCards)、在线人类孟德尔遗传数据库(OMIM)获取11个中医症状和DPN的相关靶点。利用STRING平台获取蛋白互作信息,DAVID 6.8数据库对交集靶点进行富集分析,随后基于分子对接验证关键活性成分与核心靶标的结合活性。通过蛋白质印迹法(Western Blot)检测小鼠坐骨神经组织中IL-6、ERK1/2、P38MAPK、PI3K以及AKT蛋白的表达,验证网络药理学对分子机制的预测。结果 木丹颗粒治疗气虚血瘀证DPN的主要活性成分有槲皮素、山柰酚、木犀草素等,PPI网络显示核心靶点有AKT1、ALB、IL-6、MAPK1等,“中药-靶点-通路”网络图显示木丹颗粒主要通过作用于AKT1(PI3K/AKT)、MAPK1(ERK1/2)、BCL-2(P38)、Bax(P38)、IL-6等靶点,调控MAPK等信号通路发挥治疗DPN的作用。分子对接结果证实,木丹颗粒关键成分与核心靶点有较好的亲和力,其中ALB和MAPK1与活性成分的结合活性最高。Western Blot结果显示,木丹颗粒可以显著下调DPN小鼠坐骨神经组织中IL-6、ERK1/2、P38MAPK的表达(P<0.01),显著上调PI3K、AKT的表达(P<0.01)。结论 木丹颗粒可通过多种活性成分、多个关键靶点及多种作用途径治疗气虚血瘀证DPN,主要包括抑制炎性细胞因子的表达、抑制P38 MAPK和ERK/MAPK信号通路,激活PI3K/AKT信号通路等。
Objective To decipher the molecular mechanism of Mudan Granules in treating diabetic peripheral neuropathy(DPN)with qi deficiency and blood stasis syndrome via network pharmacology method based on the principle of combination of disease with syndrome, and then verify the predicted mechanism by molecular docking and animal experiments.Methods The main active components and their targets of Mudan Granules were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Traditional Chinese Medicine Integrated Database(TCMID).Modern Medical(MM)symptoms corresponding to the 11 traditional Chinese medicine(TCM)symptoms of qi deficiency and blood stasis syndrome were collected from SymMap.The targets of the 11 TCM symtoms and DPN were acquired from GeneCards and Online Mendelian Inheritance in Man(OMIM).Protein-protein interaction(PPI)information was obtained from STRING.Enrichment analysis for the common targets shared by Mudan Granules and DPN were conducted via DAVID 6.8.Molecular docking was then carried out to verify the binding activity of the key active components to the core targets.The expression levels of IL-6,ERK1/2,P38 MAPK,PI3 K and AKT in mouse sciatic nerve tissue were measured by Western blot to verify the predicted mechanism of action.Results The main active components of MudanGranules for treating DPN with qi deficiency and blood stasis syndrome included quercetin, kaempferol, and luteolin.The PPI network showed that the core targets were AKT1,ALB,IL-6,MAPK1,etc.The“herb-target-pathway”network illustrated that Mudan Granules mainly acted on AKT1(PI3 K/AKT),MAPK1(ERK1/2),BCL-2(P38)Bax(P38),and IL-6 to regulate MAPK and other signaling pathways for treating DPN.The molecular docking results confirmed that the key components of Mudan Granules had good affinity to the core targets, and ALB and MAPK1 had the highest binding activity to the active components.Western Blot demonstrated that Mudan Granules down-regulated the expression of IL-6,ERK1/2 and P38 MAPK
作者
杜阳
于清源
任钧国
DU Yang;YU Qing-yuan;REN Jun-guo(Xiyuan Clinical Medical College,Beijing University of Chinese Medicine,Beijing100029;Beijing Key Laboratory of Pharmacology of Chinese Materia Medica,Xiyuan Hospital,Institute of Basic Medical Sciences,China Academy of Chinese Medical Sciences,Beijing100091)
出处
《世界中西医结合杂志》
2022年第3期509-518,共10页
World Journal of Integrated Traditional and Western Medicine
基金
国家重大科技专项“重大新药研发”项目(2017ZX09301018)
中国中医科学院自选课题(ZZ11-027)
中国中医科学院科技创新工程课题(CI2021A04601)。
关键词
网络药理学
分子对接
病证结合
气虚血瘀证
糖尿病周围神经病变
MAPK信号通路
木丹颗粒
Network Pharmacology
Molecular Docking
Combination of Disease with Syndrome
Qi Deficiency and Blood Stasis Syndrome
Diabetic Peripheral Neuropathy
MAPK Signaling Pathway
Mudand Granules
