Wang Yu-Guo, Dou Yong-Qi, Yan Zi-Qiao, Gao Yue
the Sixth Medical Center of PLA General Hospital Beijing 100048, China.
PLA Medical College Beijing 100853, China.
Zhongguo Zhong Yao Za Zhi. 2023 May;48(10):2810-2819. doi: 10.19540/j.cnki.cjcmm.20221223.401.
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, ~(60)Co-γ irradiation was performed for mo-deling 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, 4 941 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, etc. 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.
通过网络药理学、分子对接和细胞实验，本研究探索并验证了人参皂苷Rg_1（Rg_1）抗放射性肠炎的潜在分子机制。从中药系统药理学数据库与分析平台（BATMAN-TCM）、瑞士靶点预测数据库（SwissTargetPrediction）和基因卡片数据库（GeneCards）中检索Rg_1和放射性肠炎的靶点。利用Cytoscape 3.7.2软件和STRING数据库构建共同靶点的蛋白质-蛋白质相互作用（PPI）网络，并筛选核心靶点。采用DAVID数据库进行基因本体论（GO）术语和京都基因与基因组百科全书（KEGG）通路富集分析，以预测可能的作用机制，随后进行Rg_1与核心靶点的分子对接和细胞实验。细胞实验方面，采用~(60)Co-γ射线照射建立IEC-6细胞模型，然后用Rg_1、蛋白激酶B（AKT）抑制剂LY294002及其他药物处理，以验证Rg_1的作用效果和机制。结果显示，筛选出Rg_1潜在靶点29个、疾病靶点4941个、共同靶点25个。根据PPI网络，核心靶点有AKT1、血管内皮生长因子A（VEGFA）、热休克蛋白90α家族A类成员1（HSP90AA1）、Bcl-2样蛋白1（BCL2L1）、雌激素受体1（ESR1）等。共同靶点主要参与RNA聚合酶Ⅱ启动子转录的正调控、信号转导、细胞增殖的正调控等生物过程。KEGG通路富集分析排名前10的通路包括磷酸肌醇3-激酶（PI3K）/AKT通路、RAS通路、丝裂原活化蛋白激酶（MAPK）通路、Ras相关蛋白1（RAP1）通路和钙通路等。分子对接显示，Rg_1与AKT1、VEGFA、HSP90AA1等核心靶点具有较高的结合亲和力。细胞实验表明，Rg_1能有效提高细胞活力和存活率，降低照射后细胞凋亡率，促进AKT1和B细胞淋巴瘤-特大（BCL-XL）的表达，抑制促凋亡蛋白Bcl-2相关X蛋白（BAX）的表达。综上所述，本研究通过网络药理学、分子对接和细胞实验，验证了Rg_1减轻放射性肠炎损伤的能力。其机制是通过调节PI3K/AKT通路，从而抑制细胞凋亡。
