基于网络药理学探究黄诺玛苷对非酒精性脂肪肝的降脂作用  

Lipid-lowering effect of flavanomarein on non-alcoholic fatty liver disease based on network pharmacology

作　　者：魏晓丽 郭艳丽 张永威 冉峥 兰怡[2] 王丽凤 毛新民[1,4] WEI Xiaoli;GUO Yanli;ZHANG Yongwei;RAN Zheng;LAN Yi;WANG Iifeng;MAO Xinmin(Department of Pharmacology,School of Pharmacy,State Key Laboratory of Pathogenesis and Prevention of High Incidence in Central Asia and Peak Discipline of Integrated Traditional Chinese and Western Medicine in the Autonomous Region,Urumqi,Xinjiang Uygur Autonomous Region,830000,China;Second Department of Cadre Ward,the Fourth Affiliated Hospital,Urumqi,Xinjiang Uygur Autonomous Region,830000,China;Department of Physiology,College of Basic Medicine,Urumqi,Xinjiang Uygur Autonomous Region,830000,China;College of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi,Xinjiang Uygur Autonomous Region,830000,China)

机构地区：[1]新疆医科大学药学院药理学教研室,中亚高发病成因与防治国家重点实验室和自治区中西医结合高峰学科,乌鲁木齐830000 [2]新疆医科大学第四附属医院干部二科,乌鲁木齐830000 [3]新疆医科大学基础医学院生理学教研室,乌鲁木齐830000 [4]新疆医科大学中医学院,乌鲁木齐830000

出　　处：《第三军医大学学报》2021年第5期383-394,共12页Journal of Third Military Medical University

基　　金：国家自然科学基金地区项目(81460636);国家自然科学基金联合基金项目(U1303233)。

摘　　要：目的利用网络药理学探索黄诺玛苷抗非酒精性脂肪肝病(non-alcoholic fatty liver disease,NAFLD)的作用靶点和信号通路,研究黄诺玛苷治疗NAFLD的作用机理。方法(1)在Swiss Target Prediction数据库中检索黄诺玛苷活性靶点,DisGeNET数据库中搜索疾病靶点,通过化合物靶点与疾病靶点作Venn分析得到黄诺玛苷调节NAFLD脂代谢的预测靶点。借助Biso Genet软件构建蛋白-蛋白相互作用(protein-protein interaction,PPI)网络图,利用Metascape数据库对核心靶点进行GO功能富集与KEGG通路富集分析,用Cytoscape软件绘制"靶点-通路"网络图。(2)采用db/db小鼠模型进行体内实验验证,30只雄性db/db小鼠按随机数字表法分为3组:db/m组(正常)、db/db模型组、db/db+黄诺玛苷组(50 mg/kg)。给药12周后处死,取小鼠肝脏,检测肝脏TG、GSH、GSSG含量,HE和油红O染色观察肝脏病理学变化,qRT-PCR和Western blot检测小鼠肝脏脂代谢的关键mRNA及蛋白的表达水平。(3)HepG2细胞在游离脂肪酸(free fatty acids,FFAs,油酸∶棕榈酸=2∶1)干预成模后,分为正常组、FFAs处理组(500μmol/L)、FFAs+黄诺玛苷低中高浓度组(25、50、100μmol/L),作用24 h。油红O染色检测HepG2细胞内TG的含量。qRT-PCR和Western blot检测HepG2细胞脂代谢的关键mRNA及蛋白的表达水平。结果(1)通过Venn分析,得到黄诺玛苷对接NAFLD的29个核心靶点,主要包括过氧化物酶体增殖物激活受体(PPARγ)、血管内皮生长因子(VEGF)和丝裂原活化蛋白激酶14(MAPK14)等。GO功能富集了6个分子功能、20个生物过程、5个细胞组分。KEGG富集的关键信号通路有40条,主要包括AMPK脂代谢通路、胰岛素抵抗(IR)通路、细胞凋亡通路等。(2)动物实验结果表明,黄诺玛苷能降低db/db模型组TG水平(P<0.05),提高GSH/GSSG比例(P<0.05)。HE和油红O染色显示黄诺玛苷可逆转肝细胞的肿大,减少肝细胞内的脂滴浸润。qRT-PCR结果表明黄诺玛苷可以降低脂质Objective To explore the targets and signaling pathways of flavanomarein in the treatment of non-alcoholic fatty liver disease(NAFLD)by network pharmacology,and to investigate the effect and mechanisms in the treatment.Methods(1)The potential active targets of flavanomarein were retrieved in the Swiss Target Prediction database,and the disease-related targets were searched in the DisGeNET database.The prediction targets of lipid metabolism of NAFLD regulated by flavanomarein was obtained through Venn analysis.The Biso Genet database was used to construct the protein-protein interaction(PPI)network diagram,the Metascape database was used to analyze GO functional enrichment and KEGG pathway enrichment,and the Cytoscape software was used to draw the"target-pathway"network diagram.(2)NAFLD db/db mice was employed for in vivo verification.Thirty male db/db mice were randomly divided into db/m group,db/db model group,and db/db+flavanomarein group(50 mg/kg).After the administration of 0.5%sodium carboxymethyl cellulose solution and flavanomarein for 12 weeks,the mice were sacrificed,and the liver contents of TG,GSH and GSSG was detected.HE staining and oil red O staining were used to observe the pathological changes of the liver.qRT-PCR and Western blotting were used to detect the mRNA and protein levels of key molecules in the lipid metabolism in the liver.(3)HepG2 cells were treated with free fatty acids(FFAs,oleic acid∶palmitic acid=2∶1)and divided into normal group,FFAs treatment group(500μmol/L),and FFAs+low,medium and high concentrations of flavanomarein(25,50,100μmol/L)for 24 h.Oil red O staining was used to detect the TG content in HepG2 cells.qRT-PCR and Western blotting were used to detect the mRNA and protein levels of key molecules in the lipid metabolism of HepG2 cells.Results(1)A total of 29 core targets of flavanomarein docking NAFLD were screened through Venn analysis,including peroxisome proliferator-activated receptor-gamma(PPARγ),vascular endothelial growth factor(VEGF),and mitogen-activated pr

关 键 词：黄诺玛苷 网络药理学 非酒精性脂肪肝病 PPARγ/SREBP/FASN信号通路 降脂 

分 类 号：R282.71[医药卫生—中药学] R285.5[医药卫生—中医学]