基于AGEs-RAGE信号转导通路的鬼箭羽干预糖尿病肾病小鼠作用机制研究  

Study on mechanism of Euonymus alatus intervention in diabetic kidney disease mice based on AGEs-RAGE signal transduction pathway

作　　者：王瑾瑾[1] 牛钰琪 马倩倩 左铭钰 闫国立[1] WANG Jinjin;NIU Yuqi;MA Qianqian;ZUO Mingyu;YAN Guoli(School of Medicine,Henan University of Chinese Medicine,Zhengzhou,Henan Province 450046,China)

机构地区：[1]河南中医药大学医学院,河南省郑州450046

出　　处：《中国慢性病预防与控制》2025年第2期126-134,共9页Chinese Journal of Prevention and Control of Chronic Diseases

基　　金：国家自然科学基金(82104748);河南省科技攻关项目(242102311282)。

摘　　要：目的利用网络药理学方法和分子对接技术探究鬼箭羽治疗糖尿病肾病的作用机制,为鬼箭羽治疗糖尿病肾病潜在的作用机制提供理论依据。方法通过中药系统药理学数据库和分析平台(TCMSP)、UniProt数据库筛选鬼箭羽的活性成分及对应靶点;运用GeneCards、DisGeNet、OMIM及TTD数据库获取糖尿病肾病疾病靶点;利用Venny平台获取两者的交集靶点;使用STRING平台及Cytoscape软件构建PPI网络;通过DAVID数据库及微生信平台进行GO和KEGG富集分析;使用Auto Dock Tools 1.5.7软件进行分子对接。采用db/db小鼠为糖尿病肾病实验研究动物,鬼箭羽干预后检测小鼠肾功能(血肌酐、尿素氮、尿蛋白/尿肌酐),HE、PAS、Masson染色及透射电镜观察肾脏形态与结构;RT-qPCR、免疫组化及Western blot检测相关基因与核心蛋白。采用SPSS 27.0软件进行单因素方差分析。结果获取鬼箭羽与糖尿病肾病的交集靶点124个,PPI网络筛选出7个核心基因。KEGG富集到癌症通路、糖尿病并发症AGEs-RAGE信号通路、脂质与动脉粥样硬化通路、流体剪切应力与动脉粥样硬化通路等。最终确定鬼箭羽治疗糖尿病肾病的核心靶点为糖基化终末产物受体(RAGE)、核转录因子(NF-κB)、血管内皮生长因子(VEGF)、白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)。分子对接结果显示,鬼箭羽的活性成分与核心靶点具有较好的结合能力。动物实验研究结果显示,与模型组比较,鬼箭羽中高剂量组小鼠血肌酐、尿素氮、尿蛋白/肌酐降低,差异均有统计学意义(P<0.05),肾脏组织病理变化减轻。RT-qPCR及Western blot结果显示,与模型组比较,鬼箭羽中高剂量组RAGE、NF-κB、IL-6mRNA及蛋白表达降低(P<0.05)。免疫组化及Western blot结果显示,与模型组比较,鬼箭羽中高剂量组VEGF、TNF-α蛋白表达降低(P<0.05)。结论鬼箭羽能够有效改善糖尿病肾病结构与功能,其作用机制可能是经由Objective To investigate the mechanism of Euonymus alatus in the treatment of diabetic nephropathy using network pharmacology and molecular docking techniques,and provide the theory basis for potential mechanism of the Euonymus alatus action in the treatment of diabetic nephropathy.Methods The active components and corresponding targets of Euonymus alatus were screened through TCMSP and UniProt databases;GeneCards,DisGeNet,OMIM and TTD databases were used to obtain diabetic nephropathy disease targets;the Venny platform was used to obtain the intersection target of the two;the STRING database and the Cytoscape were used to build the PPI network;GO and KEGG enrichment analysis were performed through DAVID database and microcredit platform;the molecular docking was performed using the AutoDock Vina software.The db/db mice served as experimental animals of diabetic nephropathy;after Euonymus alatus intervention,the renal function(blood creatinine,urea nitrogen,proteinine/urine creatinine)was tested;HE,PAS,Masson staining and transmission electron microscopy were used to observe the kidney morphology and structure;RT-qPCR,immunohistochemical technique and Western blot assay were used to test the related genes and core proteins.One-way analysis of variance was used to analyze the data.The used software was SPSS 27.0.Results In the study,124 intersection targets of Euonymus alatus and diabetic nephropathy were obtained,the seven core genes were screened with the PPI network.KEGG enriched the cancer pathways,AGEs-RAGE signaling pathway in diabetic complications,the lipid and atherosclerosis pathway,and the fluid shear stress and atherosclerosis pathway.Finally,the core targets of Euonymus alatus treating diabetic nephropathy were RAGE,NF-κB,VEGF,IL-6,and TNF-α.Molecular docking results showed that the active components of the Euonymus alatus had a better binding ability with the core targets.The results of animal experiments showed that the blood creatinine,urea nitrogen,urine protein/creatinine significantly decrease

关 键 词：鬼箭羽 糖尿病肾病 网络药理学 AGEs-RAGE信号转导通路 

分 类 号：R259[医药卫生—中西医结合]