网络药理学结合转录组学方法探讨防己黄芪汤治疗肾病综合征作用机制  

Mechanism of Fangji Huangqi Tang against Nephrotic Syndrome Based on Network Pharmacology and Transcriptomics Methods

作　　者：万浩婷 刘晴 刘路瑶 谢林彤 朱慧[1,2,3] 刘晓 WAN Hao-ting;LIU Qing;LIU Lu-yao;XIE Lin-tong;ZHU Hui;LIU Xiao(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Key Laboratory of Chinese Medicine Processing,Nanjing 210023,China;Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]江苏省中药炮制重点实验室,江苏南京210023 [3]南京中医药大学国家教育部中药炮制规范化及标准化工程研究中心,江苏南京210023

出　　处：《南京中医药大学学报》2023年第4期346-358,共13页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：江苏省高等学校基础科学(自然科学)重大项目(21KJA360008)。

摘　　要：目的基于转录组学与网络药理学方法探讨防己黄芪汤治疗肾病综合征的作用机制。方法收集网络药理学和转录组学的共同靶点,进行GO/KEGG功能富集分析。构建蛋白质-蛋白质相互作用(PPI)并进行网络拓扑分析,确定潜在核心靶点。构建通路-靶点网络图,确定发挥关键作用的信号通路。采用qPCR、Western blot法和免疫荧光染色验证大鼠肾脏组织中核心靶点的mRNA和蛋白表达水平。结果PPI分析得到7个潜在核心靶点AKT1、AMPK、CPT1B、NF-κB1、P53、TGF-β1和TLR4,主要信号通路为AMPK信号通路、PI3K-Akt信号通路、PPAR信号通路、NF-κB信号通路、TGF-β信号通路、P53信号通路、MAPK信号通路、JAK-STAT信号通路和FoxO信号通路。经动物体内实验验证,防己黄芪汤显著下调AKT1、CPT1B、NF-κB1、P53、TGF-β1、TLR4的mRNA和蛋白表达水平(P<0.05),上调AMPK的mRNA和蛋白表达水平(P<0.05)。结论从网络药理学和转录组学角度初步阐明了防己黄芪汤治疗肾病综合征多成分、多靶点、多途径的整体调节特点,可为后续的药理学研究和临床应用提供依据与参考。OBJECTIVE To explore the mechanism of Fangji Huangqi Tang(FHT)in the treatment of nephrotic syndrome(NS)based on transcriptomics and network pharmacology.METHODS Intersection target genes of network pharmacology and transcriptomics were collected for GO/KEGG functional enrichment analysis.Protein-protein interactions(PPI)were constructed and network topology analysis was performed to identify potential core targets.The pathway-target network map was constructed to identify the key signaling pathways.Finally,real-time quantitative PCR(qPCR),western blot and immunofluorescence staining were used to verify the mRNA and protein expression levels of core targets in kidney tissues of rats.RESULTS Seven potential core targets AKT1,AMPK,CPT1B,NF-κB1,P53,TGF-β1 and TLR4 were identified by PPI analysis.The main signaling pathways were AMPK,PI3K-Akt,PPAR,NF-κB,TGF-β,P53,MAPK,JAK-STAT and FoxO.In animal experiments,FHT significantly down-regulated the mRNA and protein expression levels of AKT1,CPT1B,NF-κB1,P53,TGF-β1,TLR4(P<0.05),and up-regulated the mRNA and protein expression levels of AMPK(P<0.05).CONCLUSION From the perspective of network pharmacology and transcriptomics,the overall regulation features of multi-component,multi-target and multi-pathway of FHT against NS were preliminaries elaborated,which could provide the basis and reference for subsequent pharmacological research and clinical application of FHT.

关 键 词：防己黄芪汤 肾病综合征 转录组学 网络药理学 作用机制 

分 类 号：R285.5[医药卫生—中药学]