藤梨根通过PI3K/AKT信号通路治疗肝癌的网络药理学研究和实验验证

Network pharmacology and experimental validation of the PI3K/AKT signaling pathway in the treatment of hepatocellular carcinoma by Actinidia Chinensis Planch Radix

目的通过网络药理学和体外实验的方法,探讨藤梨根治疗肝细胞癌的有效成分和可能机制。方法使用药物数据库筛选出藤梨根的主要成分和靶点蛋白,利用疾病数据库挖掘肝细胞癌疾病靶点。筛选并标准化处理药物和疾病靶点,使用FunRich 3.1.3软件计算交叉核心基因,绘制韦恩图;将核心基因导入Cytoscape 3.8.2软件构建靶点网络图,通过STRING 11.0数据库构建蛋白质互作网络,通过Metascape数据库进行富集分析。对主要成分和关键靶点,利用Swiss Dock平台进行分子对接。最后通过MTT实验、侵袭实验、划痕实验和Western blot实验验证藤梨根对肝癌细胞的抑制作用。结果本研究筛选出藤梨根治疗肝细胞癌的主要有效成分有4种:槲皮素、β-谷甾醇、芦荟大黄素、儿茶素;作用靶点86个;蛋白质互作网络中主要的基因是TP53、AKT1、MAPK1、JUN及RELA等。藤梨根治疗HCC可能的生物过程是:凋亡信号通路、对无机物质的应答等。参与的主要信号通路有:肿瘤信号通路、乙肝相关通路等。各主要成分与AKT1靶点表现出极佳的对接活性。细胞实验证实了藤梨根水提取物可以抑制肝癌细胞的增殖、侵袭、迁移能力,并激活PI3K/AKT信号通路。结论本研究初步揭示了藤梨根通过多途径、多靶点作用肝细胞癌,其中PI3K/AKT信号通路是可能的机制。

Objective To explore the effective components and possible mechanisms of Actinidia Chinensis Planch Radix(ACPR)in the treatment of hepatocellular carcinoma(HCC)by network pharmacology and in vitro cell experiment.Methods The main components and target proteins of ACPR were searched by drug database,and the disease targets of HCC were mined by disease database.Drug and disease targets were screened and standardized.Funrich3.1.3 software was used to calculate core genes and draw Venn plots.The core genes were imported into Cytoscape3.8.2 software to construct the target network map.The protein-protein interaction(PPI)network was constructed through String 11.0 database.Metascape database was used for enrichment analysis.The Swissdock platform was used for molecular docking of the main components and key targets.Finally,the inhibitory effect of ACPR on HCC cells was verified by MTT,invasion,scratch and Western blot assay.Results There were four main active components(quercetin,beta-sitosterol,aloe-emodin,catechin)and 86 targets of ACPR in the treatment of HCC.The improtant genes in the PPI network were TP53,AKT1,MAPK1,JUN,RELA,etc.The possible biological processes of ACPR in the treatment of HCC were apoptosis signaling pathway,response to inorganic substances and so on.The main signaling pathways involved were tumor signaling pathway,hepatitis B-related pathway,etc.All the major components showed good binding activity with the key targets,especially-sitosterol and MAPK1.Cell experiments confirmed that ACPR could inhibit the proliferation,invade and migration of HCC cells,and activate the PI3 K/Akt signaling pathway.Conclusions The studies confirm that the mechanism of ACPR therapy for HCC involves multiple pathways and targets,and PI3 K/AKT signaling pathway is a possible mechanism.