基于网络药理学和实验验证探讨降脂理肝汤治疗非酒精性脂肪肝的作用机制  

Exploring action mechanism of Jiangzhi Ligan decoction intreatment of non-alcoholic fatty liver disease based onnetwork pharmacology and experimental validation

作　　者：季敬 姜昊 彭娇 孙妍华 何珊[1] JI Jing;JIANG Hao;PENG Jiao;SUN Yan-hua;HE Shan(College of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,China)

机构地区：[1]山东中医药大学药学院,山东济南250355

出　　处：《中国药理学通报》2025年第2期356-364,共9页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金青年科学基金资助项目(No 81903944);国家中医药管理局高水平中医药重点学科建设项目(No zyyzdxk-2023124);山东省中医药重点学科建设项目[鲁卫中医药科教字(2022)4号]。

摘　　要：目的基于网络药理学和分子对接技术,并结合细胞实验验证,探究降脂理肝汤(Jiangzhi Ligan decoction,JZLGD)治疗非酒精性脂肪肝(non-alcoholic fatty liver disease,NAFLD)的作用机制。方法通过网络药理学相关数据库获取JZLGD治疗NAFLD的活性成分、潜在靶点,构建“药物-成分-靶点”关系网络图,对交集基因进行GO和KEGG富集分析;并将核心成分与核心靶点进行分子对接。然后,采用棕榈酸(palmitic acid,PA)诱导HepG2细胞建立NAFLD模型,MTT检测JZLGD含药血清的细胞毒性;试剂盒检测胞内总胆固醇(total cholesterol,TC)、甘油三酯(triglycerides,TG)的含量;油红O染色检测细胞脂质变性;Western blot法检测自噬关键靶点p62和微管相关蛋白1轻链3(microtubule-associated-proteinlight-chain-3,LC3)、AMPKα和p-AMPKα蛋白的表达;设置AMPK抑制剂(compound C,CC)和自噬抑制剂氯喹(chloroquine,CQ)干预组,检测胞内TC和TG的含量,进一步验证JZLGD的药效及机制。结果筛选出JZLGD活性成分64个,涉及靶点373个,与NAFLD交集靶点109个,前4个核心靶点分别为INS、AKT1、TNF和PPARγ,KEGG分析显示,JZLGD治疗NAFLD主要富集在胰岛素抵抗、AMPK、mTOR、自噬等信号通路。分子对接结果显示,隐丹参酮、荷叶碱、槲皮素、芦荟大黄素等核心成分与核心靶点及自噬相关蛋白受体结合良好。细胞结果表明,JZLGD干预可以显著降低PA诱导的HepG2细胞胞内TG、TC含量,下调p62蛋白表达,上调LC3Ⅱ蛋白表达,AMPK磷酸化程度升高;而CC和CQ的干预下,很大程度上逆转了JZLGD改善脂质蓄积的作用。结论JZLGD调节AMPK信号通路激活自噬,从而治疗NAFLD。Aim To explore the effect mechanism of Jiangzhi Ligan decoction decoction treating non-alcoholic fatty liver disease(NAFLD),based on network pharmacology and molecular docking technology,combined with cell experiment verification.Methods The active ingredients and potential targets of JZLGD in the treatment of NAFLD were obtained from the network pharmacology-related database,and the“drug-ingredient-target”relationship network map was constructed,and the intersection genes were enriched by GO and KEGG.And the core components and the core target of molecular docking.Then,HepG2 cells were induced by palmitic acid(PA)to establish NAFLD model,and the cytotoxicity of JZLGD drug-containing serum was detected by MTT.The levels of intracellular total cholesterol(TC)and triglycerides(TG)were measured by the kit.Lipid degeneration was detected by oil red O staining.The expressions of p62,a key target of autophagy,and microtubule-associated proteinlight-chain-3(LC3),AMPKαand p-AMPKαwere detected by Western blot.AMPK inhibitor(compound C,CC)and autophagy inhibitor chloroquine(CQ)intervention groups were set up to detect intracellular TC and TG content,and further verify the efficacy and mechanism of JZLGD.Results A total of 64 active components of JZLGD were screened,involving 373 targets and 109 intersection targets with NAFLD.The first four core targets were INS,AKT1,TNF and PPARγ,respectively.KEGG analysis showed that JZLGD treatment of NAFLD mainly concentrated in insulin resistance,AMPK,mTOR,autophagy and other signaling pathways.Molecular docking results showed that cryptotanshinone,lotus leaf,quercetin,aloe emodin and other core components were well bound to core targets and autophagy related protein receptors.The results showed that JZLGD intervention could significantly decrease the contents of TG and TC in PA-induced HepG2 cells,down-regulate the expression of p62 protein,up-regulate the expression of LC3 II protein,and increase the phosphorylation degree of AMPK.However,the intervention of CC and CQ largely

关 键 词：降脂理肝汤 非酒精性脂肪肝 网络药理学 分子对接 作用机制 AMPK-自噬通路 

分 类 号：R-332[医药卫生] R289.5R319R393R575.5