目的 通過網(wǎng)絡藥理學與實驗驗證相結合的方法探究補骨脂乙素治療2型糖尿病的作用機制。方法 使用ECTM、SwissTargetPrediction、PubChem數(shù)據(jù)庫預測補骨脂乙素的作用靶點,使用GeneCards數(shù)據(jù)庫預測2型糖尿病的潛在靶點。使用Venn數(shù)據(jù)庫進行靶點交互分析,通過STRING數(shù)據(jù)庫計算蛋白相互網(wǎng)絡關系(PPI),通過Cytoscape 3.9.1插件MCODE和Cytohubba篩選核心靶點。通過DAVID數(shù)據(jù)庫進行基因本體(GO)和京都基因與基因組百科全書(KEGG)途徑的富集分析。建立胰島素抵抗的HepG2細胞模型,通過2-NBDG檢測細胞對葡萄糖攝取的能力,通過RT-PCR和Western blotting檢測網(wǎng)絡藥理學預測的磷脂肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)信號通路相關mRNA及蛋白表達。結果 補骨脂乙素和2型糖尿病共同作用的基因92個,PPI網(wǎng)絡(交互得分 ≥ 0.150)包含補骨脂乙素和2型糖尿病共同目標的86個節(jié)點和379個"邊"。在MCODE和Cytohubba的篩選結果交叉后,獲得了35個共同靶點。聚類分析表明,補骨脂乙素作用于2型糖尿病涉及PI3K/Akt信號通路等多種途徑及細胞氧化還原反應等多種細胞進程。補骨脂乙素顯著增強了胰島素抵抗細胞對葡萄糖的攝取能力(P<0.05、0.01)。與模型組相比,補骨脂乙素處理后顯著增加了胰島素受體(INS-R)、胰島素受體底物1(IRS1)、葡萄糖轉運蛋白2(GLUT2)、PI3K、Akt mRNA的表達(P<0.01、0.001)。與模型組相比,二甲雙胍組和補骨脂乙素組的INS-R、IRS1、GLUT2、Akt、PI3K、p-Akt、p-PI3K的蛋白表達顯著上調(diào)(P<0.05、0.01、0.001)。結論 補骨脂乙素可能通過PI3K/Akt信號通路增加胰島素抵抗細胞對葡萄糖的攝取能力,發(fā)揮其抗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.

R285

國家自然科學基金青年科學基金項目(31702024)；山東省自然科學基金青年基金項目(ZR2021QH305)；濱州醫(yī)學院啟動基金資助項目(BY2020KYQD24)