基于网络药理学及实验验证补骨脂乙素治疗2型糖尿病的作用机制  

Verification of mechanism of isobavachalcone in treatment of type 2 diabetes based on network pharmacology and experiment

作　　者：刘晓然 李志齐 李雅娜[1] 姜文国[1] 夏振红 LIU Xiao-ran;LI Zhi-qi;LI Ya-na;JIANG Wen-guo;XIA Zhen-hong(Binzhou Medical University,Yantai 264003,China)

机构地区：[1]滨州医学院,山东烟台264003

出　　处：《现代药物与临床》2023年第8期1881-1890,共10页Drugs & Clinic

基　　金：国家自然科学基金青年科学基金项目(31702024);山东省自然科学基金青年基金项目(ZR2021QH305);滨州医学院启动基金资助项目(BY2020KYQD24)。

摘　　要：目的通过网络药理学与实验验证相结合的方法探究补骨脂乙素治疗2型糖尿病的作用机制。方法使用ECTM、SwissTargetPrediction、PubChem数据库预测补骨脂乙素的作用靶点,使用GeneCards数据库预测2型糖尿病的潜在靶点。使用Venn数据库进行靶点交互分析,通过STRING数据库计算蛋白相互网络关系(PPI),通过Cytoscape 3.9.1插件MCODE和Cytohubba筛选核心靶点。通过DAVID数据库进行基因本体(GO)和京都基因与基因组百科全书(KEGG)途径的富集分析。建立胰岛素抵抗的HepG2细胞模型,通过2-NBDG检测细胞对葡萄糖摄取的能力,通过RT-PCR和Western blotting检测网络药理学预测的磷脂肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)信号通路相关mRNA及蛋白表达。结果补骨脂乙素和2型糖尿病共同作用的基因92个,PPI网络(交互得分≥0.150)包含补骨脂乙素和2型糖尿病共同目标的86个节点和379个"边"。在MCODE和Cytohubba的筛选结果交叉后,获得了35个共同靶点。聚类分析表明,补骨脂乙素作用于2型糖尿病涉及PI3K/Akt信号通路等多种途径及细胞氧化还原反应等多种细胞进程。补骨脂乙素显著增强了胰岛素抵抗细胞对葡萄糖的摄取能力(P<0.05、0.01)。与模型组相比,补骨脂乙素处理后显著增加了胰岛素受体(INS-R)、胰岛素受体底物1(IRS1)、葡萄糖转运蛋白2(GLUT2)、PI3K、Akt mRNA的表达(P<0.01、0.001)。与模型组相比,二甲双胍组和补骨脂乙素组的INS-R、IRS1、GLUT2、Akt、PI3K、p-Akt、p-PI3K的蛋白表达显著上调(P<0.05、0.01、0.001)。结论补骨脂乙素可能通过PI3K/Akt信号通路增加胰岛素抵抗细胞对葡萄糖的摄取能力,发挥其抗2型糖尿病作用。Objective To explore the mechanism of isobavachalcone in treatment of type 2 diabetes based on network pharmacology and experimental verification.Methods ECTM,SwissTargetPrediction,and PubChem database were used to predict the target of isobavachalcone,and GeneCards database was used to predict the potential target of type 2 diabetes.Target interaction analysis was performed using Venn database,PPI were calculated using STRING database,and core targets were screened using Cytoscape 3.9.1 plug-ins MCODE and Cytohubba.Enrichment analysis of GO and KEGG pathways was performed using the DAVID database.A model of insulin-resistant HepG2 cells was established,and the ability of the cells to take up glucose was detected by 2-NBDG,and the expression of mRNA and protein related to PI3K/Akt signaling pathway predicted by network pharmacology was detected by RT-PCR and Western Blotting.Results There were 92 genes that interacted with isobavachalcone and type 2 diabetes,and the PPI network(interaction score≥0.150)contained 86 nodes and 379"edges"that shared the goal of isobavachalcone and type 2 diabetes.After crossing the screening results of MCODE and Cytohubba,35 common targets were obtained.Cluster analysis shows that isobavachalcone acts on type 2 diabetes through multiple pathways such as the PI3K/Akt signaling pathway and cellular redox reactions.The experiment confirmed that isobavachalcone significantly enhanced the glucose uptake ability of insulin resistant cells(P<0.05,0.01).Compared with the model group,the mRNA expressions of INS-R,IRS1,GLUT2,PI3K,and Akt were significantly increased after isobavachalcone treatment(P<0.01,0.001).Compared with model group,the protein expressions of INS-R,IRS1,GLUT2,Akt,PI3K,p-Akt,and p-PI3K in metformin group and isobavachalcone group were significantly up-regulated(P<0.05,0.01,0.001).Conclusion Isobavachalcone may increase the glucose uptake of insulin resistance cells through PI3K/Akt signaling pathway and exert its anti-type 2 diabetes effect.

关 键 词：补骨脂乙素 2型糖尿病 磷脂肌醇-3-激酶/蛋白激酶B信号通路 胰岛素受体 胰岛素受体底物1 葡萄糖转运蛋白2 

分 类 号：R285[医药卫生—中药学]