苦参总黄酮改善非酒精性脂肪肝的作用机制及斑马鱼实验验证  

Mechanisms of total flavonoids from Sophora flavescens for the treatment of non-alcoholic fatty liver disease and experimental validation in zebrafish

作　　者：古玉凤 邓丙英 李倪仁 曾译萱 鲁思凡 朱晨 陈磊 刘怡[1,4,5] Gu Yufeng;Deng Bingying;Li Niren;Zeng Yixuan;Lu Sifan;Zhu Chen;Chen Lei;Liu Yi(School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510510,Guangdong Province,China;Department of Pharmacy,The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120,Guangdong Province,China;School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,Guangdong Province,China;Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics,Guangzhou 510515,Guangdong Province,China;Guangdong Basic Research Center of Excellence for Integrated Traditional and Western Medicine for Qingzhi Diseases,Guangzhou 510515,Guangdong Province,China)

机构地区：[1]南方医科大学中医药学院,广东省广州市510510 [2]广州中医药大学第二附属医院药学部,广东省广州市510120 [3]广东药科大学中药学院,广东省广州市510006 [4]广东省中药制剂重点实验室,广东省广州市510515 [5]广东省中西医结合防治情志病基础研究卓越中心,广东省广州市510515

出　　处：《中国组织工程研究》2025年第14期2969-2978,共10页Chinese Journal of Tissue Engineering Research

基　　金：广州市科技计划项目(202201011445),项目负责人:朱晨。

摘　　要：背景:苦参总黄酮具有抗炎、调节免疫、抗氧化、抗肝损伤等多种药理作用,其对非酒精性脂肪肝的治疗效果和作用机制尚不明确。目的:利用生物信息学、网络药理学方法及斑马鱼实验验证苦参总黄酮治疗非酒精性脂肪肝的作用机制。方法:构建非酒精性脂肪肝斑马鱼模型,观察苦参总黄酮治疗后斑马鱼肝脏脂质累积情况、病理形态改变,以及脂质累积、炎症基因的表达变化。通过TCMSP、Swiss Target Prediction、Bat-man数据库获取苦参总黄酮活性成分与非酒精性脂肪肝相关靶点;利用STRING数据库进行蛋白互作网络分析,GO功能富集和KEGG通路富集分析。基于GSE33814数据集,筛选出“苦参总黄酮-非酒精性脂肪肝”交集靶点的差异表达基因并运用R4.3.2软件进行相关性分析、受试者操作特征曲线分析,通过验证集GSE89632验证核心基因;RT-qPCR和Western blot实验验证核心通路相关基因和蛋白表达。结果与结论:①苦参总黄酮能够改善非酒精性脂肪肝斑马鱼肝脏脂质累积,显著抑制斑马鱼脂质和转氨酶水平的升高(P<0.05);调节炎症和脂代谢相关基因的表达;②网络药理学中获得168个共同靶点,Cytoscape拓扑分析综合筛选出排名前10的核心基因分别为HSP90AA1、STAT3、PIK3R1、MAPK1、AKT1、RXRA、PIK3CA、EGFR、JAK2、ESR1;GO和KEGG分析通路主要集中在胰岛素抵抗、脂质与动脉粥样硬化方面;生物信息分析共获得59个“苦参总黄酮-非酒精性脂肪肝”差异表达基因,受试者操作特征曲线分析及验证集验证得到6个核心靶点在健康人与非酒精性脂肪肝患者间有显著差异(P<0.01);③RT-qPCR和Western blot实验结果表明苦参总黄酮能够抑制非酒精性脂肪肝斑马鱼JAK2/STAT3信号通路的激活。以上结果表明,苦参总黄酮通过调节JAK2/STAT3信号通路减轻炎性反应,抑制脂质累积从而改善非酒精性脂肪肝。BACKGROUND:Total flavonoids from Sophora flavescens have a variety of pharmacological effects,including anti-inflammatory,immunomodulatory,antioxidant,and anti-hepatic injury,but the therapeutic effects and mechanisms in non-alcoholic fatty liver disease are not clear.OBJECTIVE:To reveal the mechanism of total flavonoids from Sophora flavescens in the treatment of non-alcoholic fatty liver disease using bioinformatics,network pharmacology and zebrafish experimental validation.METHODS:A zebrafish model of non-alcoholic fatty liver disease was constructed to observe lipid accumulation,pathomorphologic changes,and expression of inflammatory genes in the liver of zebrafish after treatment with total flavonoids from Sophora flavescens.The active ingredients of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease-related targets were obtained from TCMSP,Swiss Target Prediction,and Bat-man databases.STRING was used to perform protein-protein interaction network analysis,GO functional enrichment and KEGG pathway enrichment analysis.Based on the GSE33814 dataset,the differentially expressed genes of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease intersection targets were screened out.Correlation analysis and receiver operating characteristic curve were performed using R4.3.2 software.Core genes were verified by the validation set GSE89632.RT-qPCR and western blot assays were performed to verify the expression of core pathway-related genes and proteins.RESULTS AND CONCLUSION:(1)Total flavonoids from Sophora flavescens could improve lipid accumulation in the liver of zebrafish with non-alcoholic fatty liver disease,significantly inhibited the elevation of lipid and aminotransferase levels in zebrafish(P<0.05),and regulated the expression of genes related to inflammation and lipid metabolism.(2)A total of 168 common targets were obtained using the network pharmacology,and top 10 core genes,identified by Cytoscape topology analysis,were HSP90AA1,STAT3,PIK3R1,MAPK1,AKT1

关 键 词：苦参总黄酮 非酒精性脂肪肝 斑马鱼 网络药理学 生物信息学 JAK2 STAT3 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R285.5