基于网络药理学-分子对接及实验研究探讨肉桂醛联合姜黄素抗肝细胞癌的作用机制

Mechanism of cinnamaldehyde combined with curcumin against hepatocellular carcinoma based on network pharmacology，molecular docking and experimental study

关键词:  肉桂醛  姜黄素  网络药理学  肝癌  作用机制

Key words:  cinnamaldehyde  curcumin  network pharmacology  liver cancer  mechanism of action

摘要:

[目的] 基于网络药理学、分子对接和实验研究探讨肉桂醛联合姜黄素抗肝细胞癌的作用机制。[方法] 利用PharmMapper、SwissTarget、Target Net数据库筛选肉桂醛、姜黄素的相关作用靶点；TTD、Gene cards、OMIM筛选肝癌作用靶点；Venny软件筛选共同靶点；STRING数据库、Cytoscape 3.8.0软件构建蛋白相互作用网络（PPI）；通过DAVID数据库进行基因本体（GO）富集分析和京都基因与基因组百科全书（KEGG）富集分析，并进行分子对接。通过噻唑蓝（MTT）法、Hoechst 33258染色法、实时荧光定量逆转录聚合酶链式反应（RT-qPCR）法检测进行体外细胞实验验证。[结果] 经过筛选获取肉桂醛-姜黄素抗肝癌关键靶点，主要是RAC-α丝氨酸/苏氨酸蛋白激酶（AKT1）、表皮生长因子受体（EGFR）、基质金属蛋白酶-9（MMP9）、肿瘤坏死因子（TNF）等，可能涉及磷脂酰肌醇3-激酶（PI3K）/蛋白激酶 B（Akt）、白细胞介素（IL）-17、血管内皮生长因子（VEGF）等197条信号通路。分子对接表明，两药合用对AKT1具有较好的亲和性。选取PI3K/Akt信号通路上关键靶点进行体外实验验证，结果表明，与空白对照组比较，肉桂醛联合姜黄素能显著抑制SMMC-7721肝癌细胞存活率（P<0.01），显著升高凋亡率（P<0.05），并显著降低PI3K、AKT1、哺乳动物雷帕霉素靶蛋白（mTOR）、核因子-κB1（NF-κB1）mRNA的表达（P<0.01）。[结论] 肉桂醛联合姜黄素能有效抑制SMMC-7721肝癌细胞活性，诱导其凋亡，其抗肝癌作用机制可能与抑制PI3K/Akt信号通路有关。

Abstract:

[Objective] To study the mechanism of cinnamaldehyde combined with curcumin against hepatocellular carcinoma based on network pharmacology and molecular docking and experiments. [Methods] Pharm Mapper，Swiss Target，Target Net were used to screen the related targets of curcumin and cinnamaldehyde. TTD，Gene cards and OMIM databases were used to screen the target of liver cancer. Venny software was used to screen common targets. Enrichment analysis of gene ontology（GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） was carried out through DAVID database，and molecular docking was carried out. MTT assay，Hoechst 33258 staining，and RT-qPCR were used to detect the cells in vitro. [Results] By screening，key targets of cinnamaldehyde curcumin against liver cancer were obtained，mainly as follows：RAC-α serine/threonine protein kinase（AKT1）， epidermal growth factor receptor（EGFR），matrix metalloproteinase-9（MMP9） and tumor necrosis factor（TNF），which involved in 197 pathways including phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt） pathway，IL-17 pathway，vascular endothelial growth factor（VEGF） pathway，etc. Molecular docking showed that the combination of the two drugs had a good affinity for AKT1. The key targets of the PI3K/Akt pathway were selected and verified in vitro experiments. Cell experiments showed that Compared with the blank control group，cinnamaldehyde combined with curcumin could significantly inhibit the survival rate of SMMC-7721 HCC cells（P<0.01），significantly increase the apoptosis rate（P<0.05） and significantly decrease the mRNA expressions of PI3K，AKT1，mTOR，and NF-κB1（P<0.01）. [Conclusion] Curcumin combined with cinnamaldehyde can effectively inhibit the proliferation and induce apoptosis of SMMC-7721 hepatoma cells. Its anti-hepatoma mechanism may be related to the inhibition of the PI3K/Akt signaling pathway.