文献类型：期刊文献

中文题名：基于网络药理学及动物实验探讨景芪愈溃胶囊提高胃溃疡黏膜愈合质量的作用及机制

英文题名：Effect and mechanism of Jingqi Yukui Capsules on gastric ulcer mucosa healing quality: based on network pharmacology and animal experiment

作者：范珉珏[1];段永强[2,3];李能莲[1];杨晓轶[1];马骏[1];巩子汉[4];王道坤[1]

第一作者：范珉珏

机构：[1]甘肃中医药大学基础医学院,甘肃兰州730000;[2]宁夏医科大学,宁夏银川750004;[3]敦煌医学与转化教育部重点实验室,甘肃兰州730003;[4]中国中医科学院中医基础理论研究所,北京100700

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2022

卷号：47

期号：5

起止页码：1350

中文期刊名：中国中药杂志

外文期刊名：China Journal of Chinese Materia Medica

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；PubMed;

基金：甘肃省高等学校研究生导师科研项目(1006-09);甘肃省教育厅高等学校科研项目(2018A-044);甘肃中医药大学科创基金项目(KCYB2018-1)。

语种：中文

中文关键词：网络药理学;景芪愈溃胶囊;胃溃疡;溃疡愈合质量;炎症

外文关键词：network pharmacology;Jingqi Yukui Capsules;gastric ulcer;quality of ulcer healing;inflammation

摘要：该研究通过网络药理学预测景芪愈溃胶囊治疗胃溃疡的活性成分及作用机制并采用动物实验进一步验证部分关键靶点及信号通路,探讨景芪愈溃胶囊对胃溃疡的治疗效果及机制。首先通过BATMAN-TCM(Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine)、TCMSP(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform)下载得到景芪愈溃胶囊活性成分及作用靶点,通过GeneCards和OMIM(Online Mendelian Inheritance in Man)以“gastric ulcer”为关键词检索获取疾病靶点,将成分靶点和疾病靶点取交集,获得景芪愈溃胶囊治疗胃溃疡的潜在靶点。随后利用STRING在线数据库及Cytoscape 3.7.2软件构建靶点的PPI(protein-protein interaction)网络图并分析,利用Matescape数据库进行KEGG(Kyoto Encyclopedia of Genes and Genomes)信号通路富集分析并利用Omicshare在线分析平台进行富集通路可视化处理。最后采用改良Okabe法构建胃溃疡大鼠模型,分别使用高、中、低剂量的景芪愈溃胶囊,安胃疡胶囊以及雷贝拉唑钠肠溶胶囊进行干预,肉眼观察溃疡情况,采用HE染色观察大鼠胃组织形态学变化,采用考马斯亮蓝法检测大鼠胃组织iNOS的含量,综合评价大鼠胃溃疡的溃疡愈合质量;采用RT-qPCR及Western blot检测大鼠胃组织中部分蛋白的基因及蛋白的表达水平,进一步验证部分关键靶点及信号通路。经数据库分析获得景芪愈溃胶囊活性成分206个,作用靶点535个,胃溃疡的作用靶点1 305个,药物和疾病交集靶点166个。PPI网络分析以及KEGG通路富集分析显示,多个关键靶蛋白IL6、TNF、MAPK1、MAPK3、MAPK14等,以及前20条关键信号通路中的NF-κB signaling pathway、IL-17 signaling pathway、leukocyte transendothelial migration与炎症关系密切;动物实验选取关键蛋白p38 MAPK以及经典的炎症信号通路NF-κB signaling pathway进行进一步动物实验验证,景芪愈溃胶囊高剂量组大鼠胃溃疡的恢复情况最接近空白组,胃组织iNOS含量显著降低,p38 MAPK基因和磷酸化、NF-κB p65基因和蛋白表达水平显著降低。提示景芪愈溃胶囊可抑制关键蛋白p38 MAPK的磷酸化和NF-κB信号通路中NF-κB p65的表达以发挥抗炎的作用,并可有效提高胃溃疡大鼠的溃疡愈合质量,验证了网络药理学的部分预测结果。
This study aims to identify the active components and the mechanism of Jingqi Yukui Capsules(JQYK) in the treatment of gastric ulcer based on network pharmacology, and verify some key targets and signaling pathways through animal experiment. To be specific, first, the active components and targets of JQYK were retrieved from a Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM) and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and the targets of gastric ulcer from GeneCards and Online Mendelian Inheritance in Man(OMIM) with the search term "gastric ulcer". The common targets of the two were the potential targets of the prescription for the treatment of the di-sease. Then, protein-protein interaction(PPI) network of key targets were constructed based on STRING and Cytoscape 3.7.2, followed by Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment by matescape database and pathway visualization by Omicshare. For the animal experiment, the improved method of Okabe was used to induce gastric ulcer in rats, and the model rats were classified into the model group, JQYK high-dose(JQYK-H), medium-dose(JQYK-M), and low-dose(JQYK-L) groups, Anweiyang Capsules(WYA) group, and Rabeprazole Sodium Enteric Capsules(RBPZ) group. Normal rats were included in the blank group. Rats in the blank group and model group were given distilled water and those in the administration groups received corresponding drugs. Then gastric ulcer healing in rats was observed. The changes of the gastric histomorphology in rats were evaluated based on hematoxylin-eosin(HE) staining, and the content of inducible nitric oxide synthase(iNOS) in rat gastric tissue was detected with Coomassie brilliant blue method. The mRNA and protein levels of some proteins in rat gastric tissue were determined by real-time quantitative polymerase chain reaction(RT-qPCR) and Western blot(WB) to further validate some key targets and signaling pathways. A total of 206 active components and 535 targets of JQYK, 1 305 targets of gastric ulcer, and 166 common targets of the disease and the drug were yielded. According to PPI analysis and KEGG pathway enrichment analysis, multiple key targets, such as interleukin-6(IL-6), tumor necrosis factor(TNF), mitogen-activated protein kinase 1(MAPK1), MAPK3, and MAPK14, as well as nuclear factor kappa-B(NF-κB) signaling pathway, IL-17 signaling pathway, and leukocyte transendothelial migration in the top 20 key signaling pathways were closely related to inflammation. The key protein p38 MAPK and NF-κB signaling pathway were selected for further verification by animal experiment. The gastric ulcer in the JQYK-H group recovered nearly to the level in the blank group, with significant decrease in the content of iNOS in rat gastric tissue and significant reduction in the mRNA and phosphorylation levels of p38 MAPK and the mRNA and protein levels of NF-κB p65 in rat gastric tissue. The results indicated that JQYK can inhibit the phosphorylation of the key protein p38 MAPK and the expression of NF-κB p65 in the NF-κB signaling pathway, thereby exerting the anti-inflammatory effect and effectively improving the quality of gastric ulcer healing in rats. Thus, the animal experiment result verifies some predictions of network pharmacology.