摘要
目的:采用网络药理学方法和实验验证,分析土茯苓-石韦药对治疗衰老相关慢性肾脏病(aging-related chronic kidney disease,ARCKD)的有效成分、潜在作用靶点及机制,为其临床应用提供理论基础。方法:借助中药系统药理学数据库及分析平台(TCMSP)筛选该药对的有效成分,使用TCMSP、STITCH和SWISS数据库获取该药对的潜在作用靶点,并利用UniProt数据库将靶点蛋白规范化。利用The Human Phenotype Ontology、DisGeNET和Therapeutic Target Database(TTD)数据库进行疾病靶点筛选后,与药物靶点交集获取ARCKD的潜在作用靶点。通过Cytoscape软件构建“药物-活性成分-靶点”网络,将潜在作用靶点导入STRING数据库再构建蛋白互相作用(PPI)网络,进行深入分析。通过DAVID平台进行基因本体(GO)功能富集和京都基因与基因组百科全书(KEGG)通路富集分析,并检测马兜铃酸(AAI)模型小鼠的肾功能和病理染色,检测马兜铃酸诱导的NRK-52E细胞肿瘤蛋白P53(TP53)、细胞周期蛋白依赖性激酶抑制剂1A(P21)、α平滑肌激动蛋白(α-SMA)、波形蛋白(Vimentin)的表达。结果:土茯苓-石韦药对治疗衰老相关慢性肾脏病的有效成分有17个,潜在作用靶点112个,PPI靶点筛选分析获得核心靶点6个,重要靶点27个,次要靶点36个,构成以TP53为中心的复杂作用网络。GO功能富集涉及532个条目,主要影响衰老、氧化还原、炎症应答、细胞增殖的正性调节等过程;KEGG通路富集获得137条,涉及P53、癌症、NF-κB和AMPK等相关通路。动物实验结果表明,与模型对照组相比,土茯苓-石韦药对及槲皮素能改善小鼠的血肌酐、尿素氮、尿酸、尿蛋白及病理损伤;细胞实验结果证实,该药对的共有活性成分(槲皮素)能改善马兜铃酸诱导的细胞衰老与损伤,下调P53、P21、α-SMA、Vimentin蛋白的表达。结论:土茯苓-石韦药对以TP53为核心的复杂网络调节发挥肾脏保护作用,槲皮素是其关键活性成分。该药对和槲皮素能改善AAI模型小鼠肾功能以及肾脏病理损伤,其中槲皮素可有效改善AAI诱导的细胞衰老与损伤,其机制可能与下调P53信号通路有密切关系。
Objective:This study aims to analyze the effective ingredients,potential targets,and mechanism of Tufuling(土茯苓)-Shiwei(石韦)pair(hereinafter referred to as the pair)in treating the aging-related chronic kidney disease(ARCKD)based on network pharmacology and experiments,which is expected to lay a theoretical basis for the clinical application.Methods:The active ingredients of the pair were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and the potential targets from TCMSP,STITCH,and SWISS.Then the names of the target proteins were converted into the ID names of corresponding genes with UniProt.Targets related to ARCKD were searched from The Human Phenotype Ontology,DisGeNET,and Therapeutic Target Database(TTD),and the common targets of the pair and the disease were the potential targets for the treatment of the disease.The medicinal-active ingredient-target network was constructed by Cytoscape and then potential targets were imported into STRING for establishing the protein-protein interaction(PPI)network.Gene ontology(GO)term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were performed on the DAVID platform.The renal functions of aristolochic acidⅠ(AAⅠ)-induced ARCKD mice were detected and the pathological sections were observed after staining.In addition,the expression of tumor protein P53(TP53),cyclin-dependent kinase inhibitor 1 A(P21),α-smooth muscle actin(α-SMA),and vimentin in AAI-induced NRK-52 E cells was examined.Results:A total of 17 active ingredients and 112 potential targets of the pair for the treatment of ARCKD were retrieved.After screening,a PPI network involving 69 potential targets with TP53 as a key was established:6 core targets,27 important targets,and 36 secondary targets.The targets were related to 532 GO terms(mainly aging,redox,inflammatory response,and positive regulation of cell proliferation)and 137 KEGG pathways(P53,cancer,NF-κB,AMPK,etc.).Animal experiment showed that the pair and quercetin can improve serum creatinine,urea nitrogen,uric acid,and pathological injury in mice.Cell experiment results confirmed that the common active ingredient(quercetin)of the pair can alleviate AAⅠ-induced cell senescence and damage and down-regulate the expression of P53,P21,α-SMA,and vimentin.Conclusion:The pair,with the main ingredient of quercetin,protects the kidney based on the complex network with TP53 as the core.The pair and quercetin can improve renal function and pathological injury of the kidney in AAⅠ-induced ARCKD mice,and quercetin can effectively alleviate AAⅠ-induced cell senescence and damage,which may be related to the down-regulation of P53 signaling pathway.
作者
李志强
朱婧婧
周熙谋
王昉
何伟明
Li Zhiqiang;Zhu Jingjing;Zhou Ximou;Wang Fang;He Weiming(Jiangsu Province Hospital of Chinese Medicine,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029;Nanjing University of Chinese Medicine,Nanjing 210046;Medical Research Center,First College of Clinical Medicine,Nanjing University of Chinese Medicine,Nanjing 210046)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第2期146-152,共7页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金(编号:81774269)
江苏省研究生科研与实践创新计划项目(编号:KYCX20_1527)
关键词
土茯苓
石韦
网络药理学
衰老
慢性肾脏病
Tufuling
Shiwei
Network pharmacology
Aging
Chronic kidney disease
