石斛碱通过上调HSP90AA1调节免疫细胞干预糖尿病肾病  

Dendrobine interferes with diabetic kidney disease by up-regulating HSP90AA1 to regulate immune cells

作　　者：姜运会 唐传铁 李军 张楠楠 Jiang Yunhui;Tang Chuantie;Li Jun;Zhang Nannan(Shool of Basic Medicine,Guizhou University of Traditional Chinese Medicine,Guiyang 550025;Dept of Traditional Chinese Medicine,People's Hospital of Zoucheng,Jining 273500)

机构地区：[1]贵州中医药大学基础医学院,贵阳550025 [2]邹城市人民医院中医科,济宁273500

出　　处：《安徽医科大学学报》2024年第9期1599-1609,共11页Acta Universitatis Medicinalis Anhui

基　　金：2023年贵州省国家级大学生创新创业训练计划项目(编号:202310662125);贵州省科技计划项目(编号:黔科合基础-ZK[2023]一般428);贵州中医药大学大学生创新创业训练计划项目[编号:贵中医大创合字(2023)30号];贵州中医药大学博士启动基金(编号:[2021]10号)。

摘　　要：目的运用网络药理学、生物信息学和Western blot探究石斛碱通过干预免疫治疗糖尿病肾病的作用机制。方法通过Pharmmaper、Superpred等数据库筛选石斛碱的靶点,从GeneCards、OMIM等数据库筛选糖尿病肾病的疾病靶点,在Immport、InnateDB等数据库中检索免疫相关靶点,韦恩图获取上述三者之间的交集;STRING软件结合Cytoscape软件构建交集靶点的PPI图,使用CytoHubba插件挖掘互作网络中的核心靶点;从GEO获取数据集GSE142025,使用R语言CIBERSORT包进行免疫浸润和WGCNA分析;使用GSE1009、GSE30122和GSE142025进行基因差异表达分析和ROC验证,将关键靶点与免疫细胞表型进行相关性分析,并进行分子对接和分子动力学模拟分析;使用HK-2细胞进行Western blot实验检测HSP90AA1的表达。结果共得到HSP90AA1、LCK、EGFR等128个交集基因,核心靶点涉及HSP90AA1、LCK和STAT3;分子对接及分子动力学模拟分析显示HSP90AA1的蛋白可以与石斛碱进行良好的结合;Western blot实验表明:与对照组相比,高糖组中HSP90AA1显著下调;与高糖组相比,给药组中HSP90AA1显著上调。结论石斛碱可以通过调节HSP90AA1的表达从而干预糖尿病肾病。Objective To explore the mechanism of dendrobine in the treatment of diabetic kidney disease(DKD)by intervening immunity using network pharmacology,bioinformatics and Western blot.Methods The targets of dendrobine were screened by Pharmmaper,Superpred and other databases.The disease targets of DKD were screened from GeneCards,OMIM and other databases.The immune-related targets were searched in Immport,InnateDB and other databases.The intersection of the above three was obtained via Venn diagram.The PPI map of intersection targets was constructed by STRING software combined with Cytoscape software,and the core targets in the interaction network were mined by CytoHubba plug-in.The data set GSE142025 was obtained from GEO,and immune infiltration and WGCNA analysis were performed using the R language CIBERSORT package.GSE1009,GSE30122 and GSE142025 were used for gene differential expression analysis and ROC verification.The correlation between core targets and immune cell phenotype was analyzed.Molecular docking and molecular dynamics simulation analysis were used to test the binding force of dendrobine to key targets.Western blot was used to detect the expression of HSP90AA1 in HK-2 cells.Results A total of 128 intersecting genes,including HSP90AA1,LCK and EGFR,were obtained,and the core targets involved HSP90AA1,LCK and STAT3;molecular docking and molecular dynamics simulation analyses showed that the protein of HSP90AA1 could bind well with dendrobine.Western blot experiments showed that compared with the control group,HSP90AA1 was significantly down-regulated in the high glucose group.Compared with the high glucose group,HSP90AA1 was significantly up-regulated in the dendrobine group.Conclusion Dendrobine can intervene in diabetic kidney disease by regulating the expression of HSP90AA1.

关 键 词：石斛碱 糖尿病肾病 HSP90AA1 免疫细胞 网络药理学 生物信息学 靶点 

分 类 号：R285[医药卫生—中药学]