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.

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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)