人参皂苷Rg_(1)治疗放射性肠炎的网络药理学与体外实验研究

Molecular mechanism of ginsenoside Rg_(1)against radiation enteritis:based on network pharmacology and in vitro experiment

运用网络药理学、分子对接技术和细胞实验探索并验证人参皂苷Rg_(1)(ginsenoside Rg_(1),Rg_(1))治疗放射性肠炎的潜在分子机制。借助BATMAN-TCM、SwissTargetPrediction、GeneCards等数据库预测Rg_(1)和放射性肠炎的相关靶点。运用Cytoscape 3.7.2软件和STRING数据库对共同靶点进行PPI网络构建和核心靶点筛选,利用DAVID数据库进行GO功能和KEGG通路富集分析,预测其可能的作用机制。对Rg_(1)与核心靶点进行分子对接,然后进行细胞实验验证。利用60Co-γ射线照射IEC-6细胞构建模型,予以Rg_(1)、AKT阻滞剂LY294002等药物干预,验证Rg_(1)的治疗效果和作用机制。结果显示,Rg_(1)潜在作用靶点29个,疾病靶点4941个,共同靶点25个;PPI网络筛选出核心靶点包括AKT1、VEGFA、HSP90AA1、BCL2L1、ESR1等,GO分析结果主要涉及RNA聚合酶Ⅱ启动子转录的正调控、信号转导和对细胞增殖的阳性调节等生物学过程,KEGG富集分析获得PI3K/AKT通路、RAS通路、MAPK通路、RAP1通路、钙通路等10条潜在作用机制。分子对接结果显示,Rg_(1)与AKT1、VEGFA、HSP90AA1等主要核心靶点均具有较好的结合能力。细胞实验表明,Rg_(1)能够有效改善照射后细胞活力,提高存活率,降低凋亡率,并促进AKT1和BCL-XL表达,抑制促凋亡蛋白BAX的表达。综上,该研究利用网络药理学、分子对接和细胞实验阐明Rg_(1)可有效减轻肠细胞辐射损伤,其作用机制与调控PI3K/AKT通路进而抑制细胞凋亡相关。

Via network pharmacology,molecular docking,and cellular experiment,this study explored and validated the potential molecular mechanism of ginsenoside Rg_(1)(Rg_(1))against radiation enteritis.Targets of Rg_(1)and radiation enteritis were retrieved from BATMAN-TCM,SwissTargetPrediction,and GeneCards.Cytoscape 3.7.2 and STRING were employed for the construction of protein-protein interaction(PPI)network for the common targets,and screening of core targets.DAVID was used for Gene Ontology(GO)term and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment to predict the possible mechanism,followed by molecular docking of Rg_(1)with core targets and cellular experiment.For the cellular experiment,60Co-γirradiation was performed for modeling of IEC-6 cells,which were then treated with Rg_(1),protein kinase B(AKT)inhibitor LY294002,and other drugs to verify the effect and mechanism of Rg_(1).The results showed that 29 potential targets of Rg_(1),4941 disease targets,and 25 common targets were screened out.According to the PPI network,the core targets were AKT1,vascular endothelial growth factor A(VEGFA),heat shock protein 90 alpha family class A member 1(HSP90AA1),Bcl-2-like protein 1(BCL2L1),estrogen receptor 1(ESR1),etc.The common targets were mainly involved in the GO terms such as positive regulation of RNA polymerase II promoter transcription,signal transduction,positive regulation of cell proliferation,and other biological processes.The top 10 KEGG pathways included phosphoinositide 3-kinase(PI3K)/AKT pathway,RAS pathway,mitogen-activated protein kinase(MAPK)pathway,Ras-proximate-1(RAP1)pathway,and calcium pathway.Molecular docking showed that Rg_(1) had high binding affinity to AKT1,VEGFA,HSP90AA1,and other core targets.Cellular experiment indicated that Rg_(1) can effectively improve cell viability and survival,decrease apoptosis after irradiation,promote the expression of AKT1 and B-cell lymphoma-extra large(BCL-XL),and inhibit the expression of the pro-apoptotic protein Bcl-2-associated X protein(BAX).In conclusion,through network pharmacology,molecular docking,and cellular experiment,this study verified the ability of Rg_(1) to reduce radiation enteritis injury.The mechanism was that it regulated PI3K/AKT pathway,thereby suppressing apoptosis.