海南医科大学学报2025，Vol.31Issue(7)：531-540,10.DOI:10.13210/j.cnki.jhmu.20241031.001
基于网络药理学和细胞实验探究健脾活血方干预AMPK和Hippo糖脂代谢信号通路治疗肝纤维化研究
Mechanism of Jianpi Huoxue Decoction in the treatment of liver fibrosis by AMPK and Hippo glycolipid metabolism signaling pathways based on network pharmacology and cellular experiments
摘要
Abstract
Objective:To explore and verify the target and mechanism of Jianpi Huoxue Decoction in the intervention of liver fibrosis.Methods:The active components of Jianpi Huoxue Decoction were obtained by searching TCMSP,and the disease tar-gets of liver fibrosis were obtained by searching GeneCards,OMIM,TTD and DrugBank databases,and then the intersection of active components and targets was used to obtain the predictive targets of Jianpi Huoxue Decoction in the treatment of liver fibro-sis.Cytoscape 3.8.0 was used to construct the'drug-component-target-disease'network,the protein-protein interaction network was constructed by STRING database,and the core network was simplified;R software was used for GO enrichment and KEGG pathway enrichment analysis,and the components with higher degree values in the network were molecularly docked with the core targets.Human stellate cells LX-2 were cultured and divided into the blank control group,model group[induced by 5 ng trans-forming growth factor-beta 1(TGF-β1)]and observation group(5 ng TGF-β1 induction+Jianpi Huoxue Decoction interven-tion).CCK-8 method was used to detect the optimal concentration of Chinese medicine intervention cells,Western blot(WB)was used to detect the expression of α-smooth muscle actin(α-SMA),tissue inhibitor of metalloproteinase-1(Timp-1),desmin,colla-gen I and AMPK and Hippo signaling pathway proteins in each group.Results:The results showed that 114 main active ingredi-ents were screened out,involving 146 gene targets,including the main active ingredients such as luteolin,quercetin,β-sitosterol,angelica,betulinic acid,ect,the core target genes including TNF,IL-6,PTGS2,TP53,MAPK1,JUN,ESR1,etc.A total of 661 GO biological processes,467 cell components,and 115 molecular functions were enriched,including 164 KEGG signaling pathways such as fluid shear stress and atherosclerosis,AMPK,TNF,and Hippo.The results of CCK-8 showed that the prolifer-ation of LX-2 cells was the best at 0.625 mg/mL and 1.25 mg/mL.The results of WB showed that compared with the control group,the expression of α-SMA,Collagen I,Desmin and Timp-1 in the model group increased.Compared with the model group,the expression of α-SMA,Collagen I,Desmin and Timp-1 in the observation group decreased(P<0.05).WB results showed that compared with the control group,the expression of p-YAP,p-TAZ and p-AMPK in the model group decreased.Compared with the model group,the expression of p-YAP,p-TAZ and p-AMPK in the observation group increased(P<0.05).Conclu-sion:The mechanism of Jianpi Huoxue Decoction in the treatment of liver fibrosis is related to the multi-component and multi-target intervention of AMPK and Hippo signaling pathways.关键词
健脾活血方/肝纤维化/网络药理学/AMPK/Hippo信号通路Key words
Jianpi Huoxue Decoction/Liver fibrosis/Network pharmacology/AMPK/Hippo signaling pathway分类
中医学引用本文复制引用
黄迪,彭辉,李子义,胡高斌,黄龙,张琦,于庆生..基于网络药理学和细胞实验探究健脾活血方干预AMPK和Hippo糖脂代谢信号通路治疗肝纤维化研究[J].海南医科大学学报,2025,31(7):531-540,10.基金项目
This study was supported by Twelfth Five Year National Clinical Key Specialty Construction Project[Spleen and Portal Hypertension Special Project,Caishe(2013)No.239] （2013）
National Famous Traditional Chinese Medicine Expert Inheritance Studio of the State Administration of Traditional Chinese Medicine[National Medical Products Office,Renjiaohan(2021)No.270] （2021）
十二五国家临床重点专科建设项目[脾脏与门静脉高压症专项,财社(2013)239号] （2013）
国家中医药管理局全国名老中医药专家传承工作室[国中医药办人教函(2021)270号] （2021）
安徽省"十四五"临床重点专科建设推荐计划[皖卫医秘(2022)105号]Recommended Plan for the Construction of Key Clinical Specialties in Anhui Province during the 14th Five Year Plan Period[Wanwei Medical Secret(2022)No.105] （2022）
