基于网络药理学探讨大补阴丸治疗IgA肾病的作用机制

Discussion on the mechanism of action of Dabuyin Pills in treating IgA nephropathy based on network pharmacology

目的基于网络药理学探讨大补阴丸治疗IgA肾病(IgAN)的作用机制,结合动物实验加以验证。方法分别以“熟地黄”“知母”等为关键词,在中药系统药理学数据库与分析平台(TCMSP)和中药分子机制的生物信息学分析工具(BATMAN-TCM)中获取其活性成分和相应的靶点蛋白,在GeneCards数据库检索“IgAN”,筛选出大补阴丸和IgAN的交集靶点靶点;运用STRING数据库构建蛋白质-蛋白质相互作用(PPI)网络;在微生信中进行基因本体(GO)分析、京都基因与基因组百科全书(KEGG)的富集分析;采用Cytoscape构建“药物-成分-靶点”网络图;运用AutoDockTools对蛋白激酶B1(Akt1)、丝裂原活化蛋白激酶1(MAPK1)、肿瘤抑制蛋白(TP53)核心靶点进行分子对接。建立IgAN模型,造模后给予大补阴丸干预,取各组小鼠肾脏、尿液及血清,测定血清尿素氮(BUN)、24小时尿蛋白定量(24h-UTP),制作并观察切片中IgA沉积程度。结果大补阴丸与IgAN交集靶点有180个;GO富集分析涉及生物过程923个条目、细胞组分81个条目和分子功能157个条目;KEGG分析筛选出前20条信号通路,白细胞介素-17(IL-17)通路、酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)通路等;分子对接结果显示大补阴丸中槲皮素与Akt1、MAPK1等关键靶点有较好的结合能力。给予IgAN小鼠大补阴丸治疗后,较模型组其24 h-UTP及血清BUN得到改善(P<0.01),肾组织系膜区IgA沉积明显减少。结论大补阴丸对IgAN具有一定的治疗作用,其机制可能与IL-17、PI3K/Akt通路有关。

Objective To explore the mechanism of action of Dabuyin Pills in treating IgA nephropathy(IgAN)based on network pharmacology,and to verify it with animal experiments.Methods Taking"Radix Rehmanniae Preparata"and"Rhizoma Anemarrhenae"as keywords,respectively,the active components and corresponding target proteins were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and the Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMAN-TCM),and"IgAN"was searched in GeneCards database to screen out the intersection targets of Dabuyin Pills and IgAN.Protein-protein interaction(PPI)network was constructed by using STRING database.Gene ontology(GO)analysis and enrichment analysis of Kyoto encyclopedia of genes and genomes(KEGG)were carried out in Bioinformatics.The network diagram of"drug-component-target"was constructed by Cytoscape.The molecular docking of protein kinase B1(Akt1),mitogen-activated protein kinase 1(MAPK1)and tumor suppressor protein(TP53)was carried out by using AutoDockTools.The model of IgAN was established,and then Dabuyin Pills was given for intervention.The kidney,urine and serum of mice in each group were collected,and the serum blood urea nitrogen(BUN)and 24-hour urine protein quantification(24h-UTP)were measured,and the slices were prepared and the IgA deposition intensity of slices was observed.Results There were 180 intersection targets between Dabuyin Pills and IgAN.GO enrichment analysis involved 923 items of biological process,81 items of cell components and 157 items of molecular function;KEGG analysis screened out the first 20 signal pathways,including interleukin-17(IL-17)pathway,phosphatidyl inositol 3-kinase/protein kinase B(PI3K/Akt)pathway,etc.The results of molecular docking showed that quercetin in Dabuyin Pills had good binding ability with key targets such as Akt1 and MAPK1.Compared with the model group,the 24h-UTP and serum BUN were improved(P<0.01),and the IgA deposition in the mesangial area of kidney tissue was significantly reduced in IgAN mice after treatment of Dabuyin Pills.Conclusion Dabuyin Pills has a certain therapeutic effect on IgAN,and its mechanism of action may be related to IL-17 and PI3K/Akt pathways.