基于网络药理学和生物信息学探讨并验证槲皮素治疗肝癌的靶点和机制

Research and Verification of Mechanism of Quercetin in the Treatment of Liver Cancer Based on Network Pharmacology and Bioinformatics

目的 采用网络药理学方法分析槲皮素治疗肝癌的相关分子生物学机制。方法 TCMSP数据库和Pubchem数据库筛选出槲皮素作用靶点,GeneCards、OMIM和TTD数据库筛选出肝癌疾病作用靶点。利用Ri386 4.0.5软件、STRING数据库和Cytoscape 3.6.2进行蛋白互作分析,利用R语言程序、GlueGO和DAVID 6.8进行GO和KEGG信号通路富集分析,利用iGEMDOCK软件和Systemsdock数据库进行分子对接来预测槲皮素对肝癌的结合作用。采用CCK8检测槲皮素对HepG2、Hep3B、Huh-7三种肝癌细胞增殖影响,ELISA法检测槲皮素干预HepG2细胞后肿瘤坏死因子(tumor necrosis factor,TNF)-α、白细胞介素(interleukin,IL)-6、IL-1β水平,Western blot和高内涵细胞成像分析系统分别检测槲皮素干预HepG2细胞前后醛糖还原酶(aldoketo reductase family 1 member B1,AKR1B1)的表达。结果 筛选出槲皮素的自身靶点299个,筛选出槲皮素作用于肝癌的靶点70个,相关的信号通路72条,分子对接结合力最强的槲皮素-肝癌靶点为AKR1B1。槲皮素降低肝癌细胞增殖,Western blot和高内涵实验结果表明,槲皮素干预的肝癌细胞数量和AKR1B1的表达降低。炎症相关因子IL-1β、IL-6、TNF-α降低。结论 槲皮素治疗肝癌具有多成分、多靶点、多通路的特点,槲皮素治疗肝癌的潜在机制可能是与AKR1B1有关。

Objective The molecular biological mechanism of quercetin in the treatment of liver cancer was analyzed by the method of network pharmacology.Methods TCMSP database and PubChem database screened the target of quercetin and gene cards,OMIM,and TTD database screened the target of liver cancer disease.Ri3864.0.5 software,string database,and Cytoscape 3.6.2 were used for protein interaction analysis,R language program,GlueGO,and David 6.8 were used for go and KEGG signal pathway enrichment analysis and iGEMDOCK software and systems dock database were used for molecular alignment to predict the binding effect of quercetin on liver cancer.CCK8 was used to detect the effect of quercetin on the proliferation of HepG2,Hep3B,and Huh-7 liver cancer cells,and ELISA was used to detect interleukin(IL)-1β、IL-6、tumor necrosis factor(TNF)-αchanges after quercetin interfered with HepG2 cells.Western blot and high connotation cell imaging analysis system wasused to detect the expression of aldoketo reductase family 1 member B1(AKR1B1)related molecules in HepG2 cells after quercetin intervention.Results 299-self-targets of quercetin were screened,70 targets of quercetin acting on liver cancer were screened,and 72 related signal pathways.The quercetin liver cancer target with the strongest molecular docking binding ability was AKR1B1.Quercetin reduced the proliferation of liver cancer cells.Western blot and high-content experiments showed that the number of liver cancer cells and the expression of AKR1B1 decreased after quercetin intervention.Inflammation related factor TNF-α、IL-6,IL-1βdecreased.Conclusion Quercetin has the characteristics of multi-component,multi-target,and multi-channel in the treatment of liver cancer.The potential mechanism of quercetin in the treatment of liver cancer may be related to AKR1B1.