桑椹抗肝纤维化作用的网络药理学分析和实验验证  

Network pharmacological analysis and experimental verification for anti-liver fibrosis effects of Mori fructus

作　　者：乔靖怡[1,2] 李汉伟[2] 陈金香 王灿[3] 宋亚刚 狄培琰 朱平生[4] 张思森 苗明三[2] QIAO Jing-yi;LI Han-wei;CHEN Jin-xiang;WANG Can;SONG Ya-gang;DI Pei-yan;ZHU Pingsheng;ZHANG Si-sen;MIAO Ming-san(The Fifth Clinical Medical College/Zhenzhou People’s Hospital,He-nan University of Chinese Medicine,Zhengzhou HE-NAN 450046,China;Academy of Chinese Medical Sciences,He-nan University of Chinese Medicine,Zhengzhou HE-NAN 450046,China;Department of Medicine,He-nan University of Chinese Medicine,Zhengzhou HE-NAN 450046,China;College of Traditional Chinese Medicine,He-nan University of Chinese Medicine,Zhengzhou HE-NAN 450046,China)

机构地区：[1]河南中医药大学第五临床医学院/郑州人民医院,河南郑州450046 [2]河南中医药大学中医药科学院,河南郑州450046 [3]河南中医药大学药学院,河南郑州450046 [4]河南中医药大学中医学院,河南郑州450046

出　　处：《中国新药与临床杂志》2024年第1期62-69,共8页Chinese Journal of New Drugs and Clinical Remedies

基　　金：国家国际合作基地[国科外函(2016)65号];河南省博士后科研资助项目(284330);河南省高校重点科研项目(21A360022,22B360008)。

摘　　要：目的利用网络药理学方法研究桑椹抗肝纤维化的主要活性成分、靶点和通路,分析其抗肝纤维化作用机制,并通过动物实验验证。方法从中药系统药理学数据库和分析平台(TCMSP)中获取桑椹的主要活性成分及相对应的作用靶点;在OMIM数据库和GeneCards数据库中进行与炎症相关的疾病靶点预测,得到炎症相关基因靶点;使用Cytoscape 3.7.2软件构建桑椹与炎症相关疾病的成分-靶点-通路网络图;使用String数据库及Cytoscape 3.7.2软件构建蛋白-蛋白相互作用(PPI)网络,并进行基因本体(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。将50只昆明种小鼠随机分为正常组,模型组和桑椹总黄酮低、中、高剂量(75、150、300 mg·kg^(-1))组。采用四氯化碳诱导小鼠肝纤维化模型,给予桑椹总黄酮干预,检测小鼠血清生化指标和炎症指标,观察肝组织病理学变化,免疫组化法检测肝组织中基质金属蛋白酶9(MMP9)的蛋白表达。结果桑椹抗炎有关的主要活性成分有花青素、槲皮素、桑色素和β-胡萝卜素,抗炎的潜在靶点有80个,PPI网络主要包括白细胞介素(IL)-6、肿瘤坏死因子(TNF)、蛋白激酶B(AKT)1、白蛋白(ALB)、血管内皮生长因子(VEGFA)、MMP9等核心靶点。涉及到的通路主要有流体剪切应力与动脉粥样硬化、癌症通路、脂质与动脉粥样硬化和Janus激酶(JAK)-信号传导及转录激活蛋白(STAT)通路等。动物实验结果显示,与正常组相比,模型组小鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平及肝组织中炎症指标IL-6、TNF-α含量以及MMP9蛋白表达均显著升高(P<0.01)。与模型组相比,桑椹总黄酮各剂量组血清中ALT、AST水平显著降低,肝组织中IL-6和TNF-α含量及MMP9蛋白表达显著降低(P<0.05),肝损伤的病变程度减轻。结论桑椹通过花青素、槲皮素等活性成分作用于IL-6、TNF-α、MMP9等靶点发挥抗肝纤AIM To explore the main active components,targets and pathways of anti-liver fibrosis of Mori fructus by network pharmacology and animal experiment,and to explore its potential mechanism of Mori fructus against liver fibrosis.METHODS The main active components and corresponding targets of Mori fructus were obtained from the traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)and the target information was converted into target gene by Uniprot database.The targets of inflammatory disease were predicted in OMIM database and GeneCards database,and the targets of inflammatory genes were obtained.The network diagram of component-target-pathway was constructed using Cytoscape 3.7.2 software.Protein protein interaction(PPI)network was constructed using the String database and Cytoscape 3.7.2 software,and Gene Ontology(GO)biological process analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed.Fifty Kunming mice were randomly divided into six groups,including normal group,model group and low,medium and high dose of total flavonoids from Mori fructus(75,150,300 mg·kg^(-1))groups.The liver fibrosis model of mice was induced by carbon tetrachloride,and the serum biochemical indexes and predicted inflammatory indexes were detected.The histopathological changes of liver were observed,and the expression of matrix metalloproteinase 9(MMP9)protein in liver tissue was detected by immunohistochemical method.RESULTS The main anti-inflammation components of Mori fructus include cyanin,quercetin,morin andβ-carotene.There were 80 potential anti-inflammatory targets,and the PPI network mainly includes interleukin(IL)-6,tumor necrosis factor(TNF),protein kinase B(AKT)1,albumin(ALB),vascular endothelial growth factor(VEGFA)and MMP9 and other core targets.The pathways involved mainly include fluid shear stress and atherosclerosis,pathways in cancer,lipid and atherosclerosis,Janus kinase/signal transducer and activator of transcription(JAK-STAT),etc.Animal

关 键 词：桑椹 炎症 免疫 肝纤维化 网络药理学 四氯化碳 

分 类 号：R975.5[医药卫生—药品]