基于分子互作网络探讨丹蒌片干预PI3K/AKT/NF-κB/TNF通路防治非酒精性脂肪肝病的机制  

Based on molecular interaction network to explore the mechanism of action of Danlou Tablets in regulating PI3K/AKT/NF-κB/TNF pathway to prevent nonalcoholic fatty liver disease

作　　者：张新[1] 陈文娜[1] 宋囡[1] 金宏飞 张琪 何信用 刘雨彤 ZHANG Xin;CHEN Wenna;SONG Nan;JIN Hongfei;ZHANG Qi;HE Xinyong;LIU Yutong(Liaoning University of Traditional Chinese Medicine,Shenyang 110000,China)

机构地区：[1]辽宁中医药大学,沈阳110000

出　　处：《中国免疫学杂志》2022年第11期1324-1332,I0001,共10页Chinese Journal of Immunology

摘　　要：目的:基于分子互作网络探讨丹蒌片治疗非酒精性脂肪肝病(NAFLD)的潜在分子作用机制。方法:通过TCMSP、PubChem和PharmMapper数据库检索丹蒌片所有活性成分及潜在作用靶点,通过DisGeNET、Gene cards及TTD数据库检索NAFLD的潜在靶点,Uniprot数据库校正去重后上传至Venny2.1软件绘制韦恩图并取其交集。同时利用Cytoscape 3.7.1软件构建药物-活性成分-靶点网络图,DAVID数据库进行GO及KEGG分析,将结果进行可视化处理。取18只C57BL/6J小鼠随机分为正常组、模型组、丹蒌片组进行体内验证,取血清检测血脂水平,Western blot检测TNF通路相关蛋白,ELISA检测TNF-α、IL-6的表达水平。结果:共得到丹蒌片的141个有效活性成分,350个潜在靶点,478个NAFLD相关靶点;Venny2.1软件筛选出丹蒌片干预NAFLD的共有靶标蛋白115个,根据P≤0.01的GO注释分析得到335个BP信息,47个MF信息,32个CC信息,93个KEGG信号通路,主要涉及炎症反应、胆固醇代谢过程、胰岛素刺激的细胞反应、TNF信号通路等;体内结果表明,与对照组相比,模型组小鼠血清LDL-C、TG和TC水平明显升高,HDL-C水平降低(P<0.05);炎症通路蛋白PI3K、p-AKT、p-IκBa蛋白表达水平降低,TRAF2、p-IKKs、NF-κB蛋白表达水平升高(P<0.05);炎症因子TNF-α、IL-6水平显著升高(P<0.05);丹蒌片治疗后,模型组小鼠血清LDL-C、TG和TC水平明显降低,HDL-C水平升高(P<0.05);炎症通路蛋白PI3K、p-AKT、p-IκBa蛋白表达水平升高,TRAF2、p-IKKs、NF-κB蛋白表达水平降低(P<0.05);炎症因子TNF-α、IL-6水平显著降低(P<0.05)。结论:丹蒌片通过多靶点协助调控氧化应激、炎症反应、脂质代谢等多条信号通路共同发挥调控作用治疗NAFLD,进一步研究发现炎症反应促进NAFLD的发生发展,为丹蒌片的临床应用及NAFLD的临床研究提供一定的参考价值。Objective:To explore the potential molecular mechanism of action of Danlou Tablets in treatment of non-alcoholic fatty liver disease(NAFLD)based on molecular interaction network.Methods:All active ingredients and potential targets of Danlou Tablets were searched by TCMSP,PubChem and PharmMapper databases,and potential targets of NAFLD were searched by DisGeNET,Gene cards and TTD databases,and the Uniprot database was used to corrected and de-weighted and uploaded to Venny2.1 software to draw Venn diagrams and take the intersection.At the same time,Cytoscape 3.7.1 software was used to construct drug-active ingredienttarget network diagrams,and DAVID database was used for GO and KEGG analysis,and the results were visualized.Eighteen C57BL/6J mice were randomly divided into normal group,model group and Psidium vulgaris tablet group for in vivo validation,and serum was taken to detect lipid level,Western blot to detect TNF pathway related proteins,ELISA to detect expression levels of TNF-αand IL-6.Results:A total of 141 active ingredients,350 potential targets and 478 NAFLD-related targets were obtained from Danlou Tablets;Venny2.1 software screened 115 common target proteins of Danlou tablets interfering with NAFLD,and 335 BP messages,47 MF messages,32 CC messages and 93 KEGG signaling according to P≤0.01 GO annotation analysis,mainly involving inflammatory response,cholesterol metabolic process,insulin-stimulated cellular response,TNF signaling pathway,etc.In vivo results showed that compared with control group,serum LDL-C,TG and TC levels in model group were significantly increased,while HDL-C level was decreased(P<0.05).Expression levels of PI3K,p-AKT and p-IκBa protein in inflammatory pathway were decreased,while expression levels of TRAF2,p-IKKS and NF-κB protein were increased(P<0.05).Levels of inflammatory factor TNF-αand IL-6 were significantly increased(P<0.05);serum LDL-C,TG and TC levels were significantly reduced,while HDL-C level was increased in model mice after treatment with Piper betel tablets

关 键 词：分子互作网络 丹蒌片 非酒精性脂肪性肝病 炎症反应 作用机制 

分 类 号：R392[医药卫生—免疫学]