运用生物信息学探讨正肝方治疗肝癌的作用机制及实验验证研究  

Mechanism of action and experimental validation study of liver cancer treatment by Zheng Gan Decoction using bioinformatics

作　　者：宋添力 王一民 刘绪 黄胜 李海霞[3] SONG Tian-li;WANG Yi-min;LIU Xu;HUANG Sheng;LI Hai-xia(Dept of Medicine,Hubei Minzu University,Enshi Hubei 445000,China;Selenium Resocerces Research and Bioapplication Hubei Key Laboratory,Enshi Hubei 445000,China;Guang′anmen Hospital,China Academy of Traditional Chinese Medicine,Beijing 100053,China)

机构地区：[1]湖北民族大学医学部,湖北恩施445000 [2]湖北民族大学硒资源研究与生物应用湖北省重点实验室,湖北恩施445000 [3]中国中医科学院广安门医院,北京100053

出　　处：《中国药理学通报》2024年第7期1383-1391,共9页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81973682);中央高水平中医医院临床研究和成果转化能力提升项目(No HLCMHPP2023045);国家自然科学基金地区基金项目(No 32160141);湖北省自然科学基金项目(No 2022CFB173);湖北省教育厅科研项目(No B2022101)。

摘　　要：目的本研究基于生物信息学预测中药复方正肝方对肝癌的作用机制及靶点,并通过实验验证正肝方对二乙基亚硝胺(diethylnitrosamine,DEN)诱导的肝癌大鼠模型的抗癌作用机制的研究。方法通过TCMSP和HERB本草组鉴数据库收集正肝方化合物成分,并通过Uniprot蛋白数据库预测正肝方的潜在作用靶点,借助OMIM、TTD和Gene Cards数据库检索肝癌疾病靶点,然后对疾病-药物的靶点进行Venny分析,随后使用String数据库绘制蛋白互作网络,利用R 4.1.3软件和Metascape数据库对核心交集靶点进行基因本体(gene ontology,GO)富集及KEGG信号通路分析,最后运用Cytoscape 3.8.2软件进行可视化处理,构建“化合物-通路-靶点-疾病”网络图,最后利用Western blot实验进行验证。结果生物信息学结果显示,正肝方与疾病的共同靶点共有171个,正肝方治疗肝癌可能是通过木犀草素、槲皮素、β-谷固醇、常春藤皂苷元等主要活性成分,通过CASP3、Bcl-2、AKT1、IL6等核心蛋白靶点,调控Hippo信号通路、PI3K-Akt信号通路等发挥抗肝癌的作用。动物实验结果显示,Western blot检测凋亡相关蛋白Caspase-3、Bcl-2、Bax表达含量,正肝方治疗组Bax和Caspase-3蛋白表达量上调,Bcl-2蛋白表达量下调。结论本研究表明正肝方可能通过调控Caspase-3、Bcl-2、Bax凋亡相关蛋白抑制肝癌细胞的生长,促进肝癌细胞的凋亡以达到抗肝癌的作用。Aim To invepredict the mechanism of action and targets of the traditional Chinese medicine compound Zheng Gan Decoction against hepatocellular carcinoma based on bioinformatics,and to experimentally verify the mechanism of anticancer action of Zheng Gan Decoction against DEN-induced hepatocellular carcinoma in a rat model.Methods The compounds of CZLF were collected from TCMSP and HERB Herbal Histology Database,and the potential targets of CZLF were predicted from Uniprot Protein Database,and the disease targets of hepatocellular carcinoma were searched with the help of OMIM,TTD,and Gene Cards databases,and then Venny analysis was performed on the targets of the disease-drugs,and then the protein-drug interactions(PI)of CZLF were mapped by String database,which was used for the study.After that,we used String database to draw protein-protein interaction(PPI)network,R 4.1.3 software and Metascape database to enrich gene ontology(GO)and analyze KEGG signaling pathway for the core intersected targets,and finally,we used Cytoscape 3.8.2 software for visualization to construct“compound-pathway-target-disease”.Lastly,Cytoscape 3.8.2 software was used to visualize and construct a“compound-pathway-target-disease”network diagram,which was finally verified by Western blot experiment.Results Bioinformatics results showed that there were 171 common targets between Zheng Gan Decoction and diseases,and Zheng Gan Decoction might exert its anticancer effect through the main active ingredients such as lignans,quercetin,β-glutosterol,ivy saponin,etc.,as well as through the core protein targets such as CASP3,BCL2,AKT1,IL6,etc.,and the modulation of the Hippo signaling pathway and PI3K-Akt signaling pathway.The results of animal experiments showed that the expression content of apoptosis-related proteins caspase-3,BCL-2 and Bax was detected by Western blot,and the expression of Bax and caspase-3 proteins was up-regulated and the expression of Bcl-2 protein was down-regulated in the treatment group of Zheng Gan Decoction.Con

关 键 词：生物信息学 正肝方 抗肿瘤 凋亡 动物实验 作用机制 

分 类 号：R-332[医药卫生] R289.5R329.25R394.2R735.7