Mechanism of <italic style="font-style: italic">Arnebia euchroma</italic> Against Melanoma： An Exploration Based on Network Pharmacology and Experimental Verification

Ying-ying KANG; Hai-yan BAO; Min LI; Fang XU; Ying YANG; Ling CHEN; Yi-ping PU; Qian QIAN; Jian-guang LI

doi:10.13422/j.cnki.syfjx.20211514

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Mechanism of Arnebia euchroma Against Melanoma： An Exploration Based on Network Pharmacology and Experimental Verification

更新时间：2022-01-04

- Mechanism of Arnebia euchroma Against Melanoma： An Exploration Based on Network Pharmacology and Experimental Verification
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 1, Pages: 204-211(2022)
- 作者机构：
  
  新疆医科大学药学院，乌鲁木齐 830011
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20211514
  CLC： R285;R965;R289;R22;R2-031;R33
- Received：31 August 2021，
  
  Published Online：29 October 2021，
  
  Published：05 January 2022
- 稿件说明：
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康莹莹,包海燕,李敏等.基于网络药理学和实验验证探讨新疆紫草治疗黑色素瘤的分子机制[J].中国实验方剂学杂志,2022,28(01):204-211.

KANG Ying-ying,BAO Hai-yan,LI Min,et al.Mechanism of Arnebia euchroma Against Melanoma： An Exploration Based on Network Pharmacology and Experimental Verification[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(01):204-211.
康莹莹,包海燕,李敏等.基于网络药理学和实验验证探讨新疆紫草治疗黑色素瘤的分子机制[J].中国实验方剂学杂志,2022,28(01):204-211. DOI： 10.13422/j.cnki.syfjx.20211514.

KANG Ying-ying,BAO Hai-yan,LI Min,et al.Mechanism of Arnebia euchroma Against Melanoma： An Exploration Based on Network Pharmacology and Experimental Verification[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(01):204-211. DOI： 10.13422/j.cnki.syfjx.20211514.

摘要

目的

通过网络药理学及分子对接技术初步预测新疆紫草治疗黑色素瘤的活性成分、作用靶点及信号通路，通过体外实验验证可能的作用机制。

方法

通过中药系统药理学数据库（TCMSP），成分靶点预测数据库（SwissTargetPrediction）以及查阅文献获取新疆紫草药理作用的活性成分及相关靶点；通过GeneCards，在线人类孟德尔遗传病数据库（OMIM），毒性与基因比较数据库（CTD）获取黑色素瘤相关靶点；使用STRING数据库构建活性成分与相关靶点的新疆紫草-黑色素瘤靶点蛋白质-蛋白质相互作用（PPI）网络图；利用Cytoscape 3.8.2软件对新疆紫草抗黑色素瘤网络节点进行筛选分析；使用DAVID 6.8数据库对交集靶点进行基因本体（GO）功能注释和京都基因与基因组百科全书（KEGG）通路富集分析。选取新疆紫草中活性成分乙酰阿卡宁和靶点通过AutoDock vina 1.1.2软件进行分子对接验证；利用体外细胞实验验证药物抗黑色素瘤的作用。

结果

预测显示，新疆紫草与黑色素瘤共同靶点271个，其中关键靶点23个，包括基质金属蛋白酶（MMP）-9，Janus激酶（JAK）2等；KEGG富集分析预测新疆紫草治疗黑色素瘤主要作用于JAK信号转导与转录激活蛋白（STAT），酪氨酸激酶受体（ErbB），血管内皮生长因子（VEGF）等信号通路；分子对接显示新疆紫草中活性成分乙酰阿卡宁与JAK2，STAT3，VEGF，MMP-9，上皮钙黏蛋白（E-cadherin）受体表现出良好的对接活性；体外蛋白免疫印迹与实时荧光定量聚合酶链式反应（Real-time PCR）结果表明，不同剂量的乙酰阿卡宁能抑制A375细胞JAK2，STAT3，VEGF，MMP-9，E-cadherin蛋白与基因的表达（

0.05）。

结论

新疆紫草治疗黑色素瘤具有多靶点、多通路的特点，其发挥治疗作用机制可能与影响JAK2，STAT3，VEGF，MMP-9，E-cadherin等关键靶点蛋白的表达，进而抑制黑色素瘤细胞侵袭和转移能力有关。该研究为新疆紫草抗肿瘤提供了实验依据。

Abstract

Objective

To preliminarily predict the active components， action targets， and signaling pathways of

Arnebia euchroma

in the treatment of melanoma based on network pharmacology and molecular docking， and to verify its possible mechanism of action in

in vitro

experiments.

Method

The active components and related targets of

A. euchroma

were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）SwissTargetPrediction and literature， and the targets related to melanoma from the GeneCards， Online Mendelian Inheritance in Man （OMIM）， and Comparative Toxicogenomics Database （CTD）. Following the construction of the protein-protein interaction （PPI） network of active components and related targets of

A. euchroma

and melanoma-related targets using STRING， Cytoscape 3.8.2 was used for screening and analyzing the nodes in the network of

A. euchroma

against melanoma. The intersections were subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis using DAVID 6.8. Acetyl alkannin， the active component in

A. euchroma

， was docked to the target by AutoDock Vina 1.1.2. The

in vitro

experiments were then carried out to verify the anti-melanoma effect of

A. euchroma

Result

A total of 271 common targets of

A. euchroma

and melanoma were harvested， among which 23 were key targets， including matrix metalloproteinase-9 （MMP-9） and Janus kinase 2 （JAK2）. As revealed by KEGG enrichment analysis，

A. euchroma

mainly acted on Janus kinase/signal transduction and activator of transcription （JAK/STAT）， tyrosine kinase receptor （ErbB）， and vascular endothelial growth factor （VEGF） signaling pathways to resist melanoma. According to molecular docking， acetyl alkannin exhibited a good docking activity with JAK2， STAT3， VEGF， MMP-9， and E-cadherin receptors. The results of Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） showed that acetyl alkannin at different doses inhibited the protein and gene expression of JAK2， STAT3， VEGF， MMP-9， and E-cadherin in A375 cells （

0.05）.

Conclusion

A. euchroma

alleviates melanoma via multiple targets and multiple pathways， and it may exert the therapeutic effects by affecting the expression of such key target proteins as JAK2， STAT3， VEGF， MMP-9， and E-cadherin and inhibiting the invasion and metastasis of melanoma cells. This study has provided an experimental basis for the treatment of tumor with

A. euchroma

关键词

Keywords

references

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Mechanism of Arnebia euchroma Against Melanoma： An Exploration Based on Network Pharmacology and Experimental Verification

DOI：10.13422/j.cnki.syfjx.20211514

摘要

Abstract

关键词

Keywords

references