基于网络药理学、分子对接及实验验证探讨霍山石斛治疗非酒精性脂肪肝的作用机制  

Mechanisms of Dendrobium huoshanense against non-alcoholic fatty liver disease by network pharmacology combined with molecular docking and experimental validation

作　　者：邓光辉 叶梦娟 蔡肖 马梦真 吕家慧 吴静[1,2] 张晓倩 邢丽花 彭代银[1,2,3,4] 王妍妍 俞年军[1,2,3,4] DENG Guang-hui;YE Meng-juan;CAI Xiao;MA Meng-zhen;LYU Jia-hui;WU Jing;ZHANG Xiao-qian;XING Li-hua;PENG Dai-yin;WANG Yan-yan;YU Nian-jun(Anhui University of Chinese Medicine,Hefei 230012,China;Anhui Academy of Traditional Chinese Medicine&Institute of Conservation and Development of Traditional Chinese Medicine Resources,Hefei 230012,China;MOE-Anhui Joint Collaborative Innovation Center for Anhui Genuine Chinese Medicinal Materials,Hefei 230012,China;Key Laboratory of Traditional Chinese Medicine Research and Development of Anhui Province,Hefei 230012,China)

机构地区：[1]安徽中医药大学,安徽合肥230012 [2]安徽省中医药科学院中药资源保护与开发研究所,安徽合肥230012 [3]省部共建安徽道地性中药材品质提升协同创新中心,安徽合肥230012 [4]中药研究与开发安徽省重点实验室,安徽合肥230012

出　　处：《中草药》2023年第16期5244-5256,共13页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金联合基金项目(U19A2009);省部共建安徽道地中药材品质提升协同创新中心项目(教科信厅函[2022]4号);安徽省教育厅项目(皖教秘科[2014]44号)。

摘　　要：目的基于网络药理学、分子对接及体内实验验证探讨霍山石斛对非酒精性脂肪肝(non-alcoholic fatty liver disease,NAFLD)的疗效及作用机制。方法利用文献检索霍山石斛活性成分,采用SwissTargetPrediction数据库预测其潜在靶点,采用DisGeNET和GeneCards数据库筛选NAFLD靶点,并将药物靶点和疾病靶点取交集,构建蛋白质-蛋白质相互作用网络,借助DAVID数据库进行基因本体功能和京都基因与基因组百科全书通路富集分析,构建“活性成分-靶点-疾病-通路”网络,并采用AutoDock等软件对关键活性成分和核心靶点进行分子对接验证。采用高脂高糖饮食诱导NAFLD小鼠模型,设置对照组、模型组、吡格列酮(10 mg/kg)组和霍山石斛低、中、高剂量(200、400、600 mg/kg)组,连续给药8周。检测血清三酰甘油(triglyceride,TG)、总胆固醇(total cholesterol,TC)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)及天冬氨酸氨基转移酶(aspartate aminotransferase,AST)的含量;ELISA检测肝脏中TC、TG、白细胞介素-6(interleukin-6,IL-6)、IL-1β及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平;油红O染色和苏木素-伊红(HE)染色检测肝组织病理变化;Western blotting检测肝组织中胰岛素受体(insulin receptor,InsR)、磷脂酰肌醇-3-激酶(phosphatidylinositol-3-kinase,PI3K)、蛋白激酶B(protein kinase B,Akt)、糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)蛋白表达。结果霍山石斛中筛选出石斛酚、铁皮石斛素A、4,4′-dihydroxy-3,5-dimethoxydihydrostilbene、柚皮素等38个活性成分,与NAFLD共同靶点有155个,作用于胰岛素、AKT1、白蛋白、TNF、IL-6、血管内皮生长因子A、过氧化物酶体增殖物激活受体γ、IL-1β等19个核心靶点,主要涉及癌症途径、PI3K/Akt通路、晚期糖基化�Objective To explore the therapeutic effect and mechanism of Dendrobium huoshanense on non-alcoholic fatty liver disease(NAFLD)based on network pharmacology,molecular docking and in vivo experimental verification.Methods The active components of D.huoshanense were searched by literature,the potential targets were predicted by SwissTargetPrediction database,the targets of NAFLD were screened by DisGeNET and GeneCards databases,drug targets and disease targets were intersected to construct protein-protein interaction network.Gene ontology function and Kyoto encyclopedia of genes and genomes pathway enrichment analysis were carried out by DAVID database,and“active ingredient-target-disease-pathway”network was constructed,molecular docking verification of key active ingredients and core targets was carried out by AutoDock and other software.NAFLD mouse model was induced by high-fat and high-sugar diet,control group,model group,pioglitazone group(10 mg/kg)and D.huoshanense low-,medium-and high-dose(200,400,600 mg/kg)groups were set up,drugs were continuously given for eight weeks.Levels of triglyceride(TG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C),alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum were detected;The levels of TC,TG,interleukin-6(IL-6),IL-1β and tumor necrosis factor-α(TNF-α)in liver were detected by ELISA.The pathological changes of liver tissue were detected by oil red O staining and hematoxylin-eosin(HE)staining.Western blotting was used to detect the expressions of insulin receptor(InsR)/phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt)/glycogen synthase kinase-3β(GSK-3β)pathway related proteins in liver tissue.Results A total of 38 active components were screened from D.huoshanense,including moscatilin,dendrobixin A,4,4′-dihydroxy-3,5-dimethohydroxystilbene,naringin,and 155 common targets with NAFLD,acting on 19 core targets such as insulin,protein kinase B1(AKT1),albumin,tumor necrosis facto

关 键 词：霍山石斛 非酒精性脂肪肝 网络药理学 芦丁 石斛酚 柚皮素 InsR/PI3K/Akt/GSK-3β通路 胰岛素抵抗 炎症 

分 类 号：R285.5[医药卫生—中药学]