基于网络药理学和实验验证探索紫铆亭治疗急性髓系白血病的机制

Exploration of the mechanism of fraxetin in treating acute myeloid leukemia based on network pharmacology and experimental verification.

作者信息

Chai Yihong, Sun Xiaohong, Zhou Qi, Li Hongxing, Xi Yaming

机构信息

The First Clinical Medical College of Lanzhou University, Lanzhou 730000, Gansu, People's Republic of China.

Department of Hematology, First Hospital of Lanzhou University, Lanzhou 730000, Gansu, People's Republic of China.

出版信息

Heliyon. 2024 Jul 16;10(15):e34717. doi: 10.1016/j.heliyon.2024.e34717. eCollection 2024 Aug 15.

DOI:10.1016/j.heliyon.2024.e34717

PMID:39166080

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11334658/

Abstract

OBJECTIVE

To explore the pharmacological mechanism of the effect of fraxetin in treating acute myeloid leukemia (AML) by the network pharmacology method combined with experimental validation.

METHODS

The targets of fraxetin were identified through Swisstarget prediction, PhammerMap, and CTDBASE. Disease-related targets of AML were explored using GeneCards and DisGenet databases, and the intersected targets were analyzed in the String website to construct a protein-protein interaction (PPI) network. Subsequently, gene ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were conducted using the DAVID database. Molecular docking of core proteins with drugs was performed using Auto Dock Vina software. Finally, the effect of fraxetin on AML was evaluated by in vitro experiments. The effect of fraxetin on AML cell proliferation was assessed by CCK8, the effect of fraxetin on AML cell apoptosis was assessed by flow cytometry, and the expression of relevant protein targets was detected by Western blotting to evaluate the anti-AML effect of fraxetin.

RESULTS

In this study, fraxetin exerts its effect against AML through 101 intersecting genes. The pathway enrichment analysis revealed that the pharmacological effects of fraxetin on AML were related to the Adenosine 5'-monophosphate (AMP)-activated protein kinase （AMPK） signaling pathway, and the molecular docking results indicated that fraxetin had an excellent binding affinity to both the core target and AMPK. In vitro experiments have demonstrated that fraxetin inhibited the proliferation and induced apoptosis of THP1 and HL60 cells, and the western blotting results indicated that the -AMPK of the fraxetin intervention group was significantly changed in a dose-dependent manner.

CONCLUSION

Fraxetin may modulate the AMPK signal pathway by interactine with the core target, thereby potentially therapeutic effect on AML.

摘要

目的

采用网络药理学方法结合实验验证，探讨紫丁香苷治疗急性髓系白血病（AML）的药理机制。

方法

通过Swisstarget预测、PhammerMap和CTDBASE鉴定紫丁香苷的靶点。利用GeneCards和DisGenet数据库探索AML的疾病相关靶点，并在String网站上分析交集靶点以构建蛋白质-蛋白质相互作用（PPI）网络。随后，使用DAVID数据库进行基因本体（GO）功能富集和京都基因与基因组百科全书（KEGG）通路富集。使用Auto Dock Vina软件对核心蛋白与药物进行分子对接。最后，通过体外实验评估紫丁香苷对AML的作用。通过CCK8评估紫丁香苷对AML细胞增殖的影响，通过流式细胞术评估紫丁香苷对AML细胞凋亡的影响，并通过蛋白质印迹法检测相关蛋白靶点的表达，以评估紫丁香苷的抗AML作用。

结果

在本研究中，紫丁香苷通过101个交集基因发挥其对AML的作用。通路富集分析表明，紫丁香苷对AML的药理作用与5'-单磷酸腺苷（AMP）激活的蛋白激酶（AMPK）信号通路有关，分子对接结果表明紫丁香苷对核心靶点和AMPK均具有良好的结合亲和力。体外实验表明，紫丁香苷抑制THP1和HL60细胞的增殖并诱导其凋亡，蛋白质印迹结果表明，紫丁香苷干预组的-AMPK以剂量依赖性方式显著变化。