基于网络药理学及动物实验探讨清热止血方治疗IgA血管炎性肾炎的机制  

Mechanism of Qingre Zhixue granules in treatment of IgA vasculitis nephritis based on network pharmacology and animal experiments

作　　者：徐闪闪[1,2] 韩姗姗[1,2] 丁樱[1,2] 代彦林 王龙[1,2] 徐炎 张霞[1,2] XU Shan-shan;HAN Shan-shan;DING Ying;DAI Yan-lin;WANG Long;XU Yan;ZHANG Xia(Pediatric Hospital,the First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,China;College of Pediatrics,Henan University of Traditional Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第一附属医院儿科医院,河南郑州450000 [2]河南中医药大学儿科医学院,河南郑州450000

出　　处：《中国药理学通报》2025年第4期762-771,共10页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金青年项目(No.82305311、82305310);河南省中医药科学研究专项(No.2023ZXZX1132);河南中医药大学“双一流”创建项目(No.HSRP-DFCTCM-2023-7-12)。

摘　　要：目的基于网络药理学及动物实验探讨清热止血方治疗IgA血管炎性肾炎(IgAVN)的作用机制。方法通过TCMSP数据库筛选出清热止血方药物的活性成分及靶点,利用Disgenet、Genecards OMIM、TTD数据库检索IgAVN疾病靶点,利用微生信在线数据库获取交集靶点,并将信息导入Cytoscope 3.7.1软件和STRING分析平台,构建药物-活性成分-治疗靶点网络;基于核心靶点通过Metascape数据库进行GO和KEGG通路富集分析,选取关键靶点及核心活性成分。通过动物实验验证网络药理学结果,采用病证结合法构建IgAVN动物模型,药物干预后检测各组IgAVN模型大鼠的尿红细胞和24 h尿蛋白,观察肾组织的HE染色和IgA免疫荧光沉积,采用Western blot和qRT-PCR检测肾组织相关蛋白的表达。结果网络药理学:获得药物活性成分109个,药物靶点402个,疾病靶点1285个,交集靶点113个。蛋白质相互作用及网络拓扑分析筛选到核心靶点是IL6、TNF、VEGFA等,核心药物成分是槲皮素、芍药苷、木犀草素、山奈酚和黄芩素等。GO富集到基因功能2545个,KEGG富集到基因通路164条,清热止血方可通过调节TNF信号通路、MAPK信号通路、IL-17信号通路、HIF-1信号通路、Toll受体信号通路、NF-κB信号通路和细胞凋亡等治疗IgAVN。动物实验:与空白组比,模型组大鼠尿红细胞和24 h尿蛋白定量升高(P<0.05);血清IL-6、TNF-α和VEGF-α表达升高(P<0.05);HE染色可见肾小球系膜增生,毛细血管的管腔狭窄,伴有少量炎性细胞浸润,肾小球有IgA免疫荧光的沉积;肾组织TNF-α、p-p65和p-p38蛋白和TNF-αmRNA表达升高(P<0.05)。清热止血方干预后尿红细胞、24 h尿蛋白定量降低(P<0.05);肾脏HE染色改变改善、肾小球IgA沉积减少;肾组织TNF-α、p-p65、p-p38蛋白和TNF-αmRNA降低(P<0.05)。结论清热止血方中槲皮素、芍药苷、木犀草素等活性成分调控TNF信号通路、MAPK信号通路、IL-17信号通路降低IL-6、TNAim To investigate the mechanism of action of Qingre Zhixue granules in the treatment of IgA vasculitic nephritis(IgAVN)based on network pharmacology and animal experiments.Methods The active ingredients and targets of Qingre Zhixue granules were screened out by TCMSP database.The disease targets of IgAVN were retrieved by Disgenet,Genecards OMIM and TTD databases.The intersection targets were obtained by WeChat online database,and the information was imported into Cytoscope 3.7.1 software and STRING analysis platform to construct a drug-active component-therapeutic target network.Based on the core targets,GO and KEGG pathway enrichment analysis was performed through the Metascape database to select key targets and core active components.The results of network pharmacology were verified by animal experiments.The animal model of IgAVN was constructed by the combination of disease and syndrome.The urine red blood cells and 24-hour urine protein of IgAVN model rats after intervention of Qingre Zhixue granules were detected.HE staining and IgA immunofluorescence deposition of renal tissue were observed.Western blot and qRT-PCR were used to detect the expression of core targets in renal tissue.Results Network pharmacology showed that 109 active ingredients,402 drug targets,1285 disease targets and 113 intersection targets were obtained.The core targets screened by protein interaction and network topology analysis were IL6,TNF,VEGFA,etc.The core drug ingredients were quercetin,paeoniflorin,luteolin,kaempferol and baicalein.A total of 2545 gene functions were enriched by GO,and 164 gene pathways were enriched by KEGG.Qingre Zhixue granules could treat IgAVN by regulating TNF signaling pathway,MAPK signaling pathway,IL-17 signaling pathway,HIF-1 signaling pathway,Toll receptor signaling pathway,NF-κB signaling pathway and apoptosis.Animal experiments showed that compared with the blank group,the urine red blood cells and 24-hour urine protein quantification in the model group increased(P<0.05);the expression of serum IL

关 键 词：清热止血方 IgA血管炎性肾炎 网络药理学 TNF-α/MAPK/NF-κB信号通路 炎症反应 机制研究 

分 类 号：R-332[医药卫生] R289.5R319R392.11R543R692.3