文献类型：期刊文献

中文题名：基于网络药理学和实验验证探讨淫羊藿苷联合泼尼松治疗激素抵抗性肾病综合征大鼠的作用机制

英文题名：To Explore the Mechanism of Icariin Combined with Prednisone on Steroid-Resistant Nephrotic Syndrome Based on Network Pharmacology and Experimental Verification

作者：吕娟[1,2];戴恩来[1];张云霞[1,2];白俊嫄[1];蒲晓薇[1]

第一作者：吕娟

机构：[1]甘肃中医药大学中西医结合学院,兰州730000;[2]甘肃省中医院,兰州730050

第一机构：甘肃中医药大学中西医结合学院

年份：2023

卷号：25

期号：7

起止页码：2426

中文期刊名：世界科学技术-中医药现代化

外文期刊名：Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD_E2023_2024】；

基金：国家自然科学基金委员会地区科学基金项目(81760799):淫羊藿苷对激素抵抗型肾病综合征大鼠GR/SD2AP及足细胞自噬的调节作用研究,负责人:戴恩来。

语种：中文

中文关键词：网络药理学;分子对接;淫羊藿苷;泼尼松;激素抵抗性肾病综合征;PI3K-Akt信号通路

外文关键词：Network pharmacology;Molecular docking;Icariin;Prednisone;Steroid-sensitive nephrotic syndrome;PI3K-Akt signaling pathway

摘要：目的基于网络药理学、分子对接以及实验验证探讨淫羊藿苷联合泼尼松治疗激素抵抗性肾病综合征(Steroid-sensitive nephrotic syndrome,SRNS)大鼠的作用机制。方法经由TCMSP、GeneCards及CTD数据库确定淫羊藿苷活性成分作用靶点;通过GeneCards基因库、OMIM遗传数据库、CTD靶点数据库以及DRUGBANK药物数据库筛选与SRNS相关的靶点,去重并整合;并绘制Venn图获取交集靶点;利用STRING数据库构建蛋白质-蛋白质互作网络,借助Cytoscape 3.7.2进行拓扑分析筛选核心靶点;使用DAVID 6.8数据库对靶点进行GO和KEGG功能富集分析。用AutoDockTools 1.5.6软件及PyMOL软件进行分子对接研究。通过构建SRNS大鼠模型,检测各组大鼠体质量、24 h尿蛋白定量、HE染色观察各大鼠肾组织病理形态;采用Western blot实验检测核心靶点蛋白表达。结果通过网络药理学分析,获得淫羊藿苷活性成分对应的靶点139个,与1476个SRNS相关靶点取交集,得到58个共同靶点,PPI网络中degree值排在前3位的分别是Akt、JUN、IL6,并且KEGG富集分析发现淫羊藿苷主要通过调控PI3K/AKT通路发挥对SRNS的治疗作用。分子对接验证显示,Akt、PI3K以及自噬相关蛋白LC3与淫羊藿苷有较好的结合活性。动物实验研究结果表明,淫羊藿苷联合泼尼松明显增加了SRNS大鼠的体质量(P<0.05)、降低了24 h尿蛋白定量(P<0.01)、改善了肾小球的形态学变化。Western blot实验结果表明,淫羊藿苷联合泼尼松可显著改善SRNS大鼠肾组织LC3-II蛋白的异常降低的Akt和PI3K蛋白表达水平的异常升高(P<0.01)。结论淫羊藿苷可通过多靶点的、多通路途径发挥治疗SRNS的作用,调节PI3K-Akt信号通路,升高SRNS大鼠肾组织自噬活性可能是其主要作用机制之一。
Objective Based on network pharmacology,molecular docking and animal experiment,this study explored the mechanism of Icariin combined with Prednisone in the treatment of steroid-resistant nephrotic syndrome(SRNS)in rats.Methods The targets of Icariin active components were determined by TCMSP,GeneCards and CTD databases.The targets of SRNS were screened from GeneCards,OMIM,CTD and DRUGBANK databases,Remove and integrate.Venn graph was drawn to obtain the intersection target.A protein-protein interaction network was constructed using STRING database,and core targets were screened by topological analysis using Cytoscape 3.7.2.DAVID 6.8 was employed for GO term enrichment and KEGG pathway enrichment.And AutoDockTools Vina,PyMOL for molecular docking.SRNS rat model was constructed.The body weight of rats was measured,The 24-hour urinary protein quantity was detected,and the pathological morphology of renal tissue was observed by HE staining.,and the expression of core target proteins was de tected by Western blot.Results Network pharmacologic analysis showed that 139 targets corresponding to Icariin active ingredients were obtained,and 58 common targets were obtained by intersection with 1476 SRNS related targets.In PPI network,the top three degree values were Akt,JUN and IL6.KEGG enrichment analysis showed that Icariin played a therapeutic role in SRNS mainly by PI3K/AKT singnaling pathway.Molecular docking verification showed that Akt,PI3K and autophagy-related protein LC3-II had good binding activity with Icariin.Animal experiment result showed that Icariin combined with Prednisone significantly increased the body weight of SRNS rats(P<0.05),decreased the 24 hour urine protein quantity(P<0.01),and improved the glomerular morphological changes.The results of Western-blotting showed that Icariin combined with Prednisone could significantly improve the abnormal decrease of LC3-II protein and the abnormal increase of Akt and PI3K protein expression in renal tissue of SRNS rats(P<0.01).Conclusion Icariin can play a role in the treatment of SRNS through a multi-target and multi-pathway,regulate the PI3K-Akt signaling pathway,and increase the autophagy activity of kidney tissue in SRNS rats,which may be one of its main mechanisms.