基于网络药理学和分子生物学探讨温下方治疗肺癌的作用机制  

Mechanisms of Warm Purgative Recipe for Treatment of Lung Cancer Based on Network Pharmacology and Molecular Biology

作　　者：周莹[1] 张韧[1] 黄意勉 廖锦彬 余泳仪 罗海燕[1] 黄少伟[1] ZHOU Ying;ZHANG Ren;HUANG Yi-Mian;LIAO Jin-Bin;YU Yong-Yi;LUO Hai-Yan;HUANG Shao-Wei(Basic Medical School,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;Guangdong Second Traditional Chinese Medicine Hospital/Guangdong Research Institute of Traditional Chinese Medicine Manuracturing Technology,Guangzhou 510095 Guangdong,China)

机构地区：[1]广州中医药大学基础医学院,广东广州510006 [2]广东省第二中医院/广东省中医药工程技术研究院,广东广州510095

出　　处：《广州中医药大学学报》2020年第10期1956-1963,共8页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：广东省科技计划项目“创新中药研发平台建设”(编号:2017A070701017);广东省中医药科技创新平台建设项目(粤中医函)。

摘　　要：【目的】应用网络药理学方法预测温下方(由人参、大黄、附子、当归组成)治疗肺癌疾病的作用靶点和信号通路,并通过细胞实验验证其预测的准确性。【方法】搜索TCMSP数据库获得温下方组成药物的有效活性成分,使用Cytoscape软件建立活性成分-作用靶点网络,使用String数据库和Cytoscape软件绘制蛋白相互作用网络,并通过DAVID数据库对靶点进行基因本体论(GO)注释及京都基因与基因组百科全书(KEGG)通路分析。制备不同浓度温下方含药血清作用A549细胞,采用四甲基偶氮唑盐(MTT)方法测定细胞增殖情况,采用定量聚合酶链反应(qPCR)技术测定苏氨酸激酶(AKT)的mRNA表达水平,验证预测结果的可靠性。【结果】温下方作用于肺癌的有效活性成分有32个,相关靶点112个,在靶点间的蛋白-蛋白相互作用网络中,分析出温下方中有39个重要靶点与治疗肺癌有关。验证实验可知,温下方含药血清能浓度依赖性抑制A549细胞的增殖,下调AKT mRNA水平。【结论】温下方治疗肺癌具有多成分-多靶点-多途径的作用特点,可为深入研究其机制指明方向。Objective To predict the targets and signaling pathways in Warm Purgative Recipe including Radix Ginseng, Radix et Rhizoma Rhei, Radix Aconiti Lateralis Preparata, Radix Angelicae Sinensis, and so on treatment for lung cancer based on network pharmacology method,and to verify the results. Methods The active ingredient of each Chinese herbs in Warm Purgative Recipe were collected from TCMSP database,and an active ingredient-target network was built by String database and Cytoscape software,and a protein-protein interaction network was established by String database and Cytoscape software. And then the achieved targets were given gene ontology(GO)note and Kyoto encyclopedia of genes and genomes(KEGG)pathway analysis by DAVID database.Furthermore, the A549 cells were cultured by different concentrations of prepared Warm Purgative Recipecontaining serum,and then the proliferative viability of A549 cells was determined by methyl thiazolyl tetrazolium(MTT)assay, and the mRNA level of AKT was determined by quantitative polymerase chain reaction(qPCR)technique. Results There were 32 chemical components,112 related targets in Warm Purgative Recipe acting on lung cancer. In protein-protein interaction network, 39 key targets related with lung cancer treatment were achieved. The verified results showed that Warm Purgative Recipe-containing serum inhibited the proliferation viability of A549 cells and down-regulated the AKT mRNA expression level in a dose-dependent manner.Conclusion Warm Purgative Recipe has multi-component, multi-target, and multi-pathway mode for the treatment of lung cancer,indicating directions to further study its mechanism.

关 键 词：温下方 肺癌 网络药理学 A549细胞 

分 类 号：R285.5[医药卫生—中药学]