目的 运用网络药理学和实验探究瑞巴派特改善慢性萎缩性胃炎小鼠的作用靶点及通路。方法 通过PubChem中进行检索得到瑞巴派特的详细信息，并分别导入Targetnet、SwissTarget Prediction、Super-PRED、PharmMapper数据库进行检索得到与瑞巴派特作用相关的靶点。通过在Pharmgkb、OMIM、GeneCards、Disgnet、CTD数据库筛选出慢性萎缩性胃炎相关的疾病靶点。利用韦恩图获取药物与疾病的交集靶点，将获得的交集靶点导入STRING数据库，建立蛋白相互作用（PPI）网络图，借助DAVID进行基因本体论（GO）功能、京都基因和基因组百科全书（KEGG）通路富集分析，最后对可能的核心靶点进行分子对接验证。采用幽门螺杆菌和亚硝酸盐建立慢性萎缩性胃炎小鼠模型，通过苏木素–伊红（HE）染色、免疫组化染色法、实时荧光定量（RT-qPCR）法验证瑞巴派特对慢性萎缩性胃炎的炎症因子和部分关键靶点的调控作用。结果 通过数据库筛选出相关药物靶点506个，疾病靶点2 115个，“药物–疾病”交集靶点169个，共获得20个核心靶点、10类生物过程，凋亡及叉头转录因子（FoxO）信号通路等20条相关通路，分子对接显示瑞巴派特与蛋白激酶B1（Akt1）蛋白、FoxO蛋白、磷脂酰肌醇-3-激酶（PI3K）蛋白亲和力良好。实验证实了瑞巴派特降低了促炎因子[白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、γ干扰素（IFN-γ）、白细胞介素-1β（IL-1β）]的mRNA表达，升高抑炎因子[白细胞介素-4（IL-4）、白细胞介素-10（IL-10）]、胃泌素和H⁺/K⁺-ATP酶的mRNA表达，并上调FoxO信号通路上的PI3K、AKT mRNA水平，下调FoxO水平。结论 瑞巴派特可能通过调控FoxO通路，调节凋亡过程、抑制炎性反应，进而有效缓解慢性萎缩性胃炎小鼠胃黏膜损伤。

Objective To investigate the action targets and pathways of rebamipide in treatment of mice with chronic atrophic gastritis by using network pharmacology and experiments. Methods The detailed information of rebamipide was obtained through retrieval in PubChem, and the target related to rebamipide action was retrieved by importing Targetnet, SwissTarget Prediction, Super-PRED, and PharmMapper database respectively. Disease targets related to chronic atrophic gastritis were selected from Pharmgkb, OMIM, GeneCards, Disgnet, and CTD databases. The intersection targets of drugs and diseases were obtained using Venn diagram, which was imported into STRING database to establish the protein interaction (PPI) network map. The gene ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were carried out with DAVID. Finally, the possible core targets were verified by molecular docking. Helicobacter pylori and nitrite were used to establish a mouse model of chronic atrophic gastritis. Hematoxylin-eosin (HE) staining, immunohistochemical staining, and real-time fluorescence quantization (RT-qPCR) were used to verify the regulatory effects of repaxide on inflammatory factors and some key targets of chronic atrophic gastritis. Results A total of 506 drug targets, 2 115 disease targets, and 169 drug-disease intersection targets were screened out through the database. A total of 20 core targets, 10 biological processes, and 20 related pathways including apoptosis and FoxO signaling pathway were obtained. Molecular docking showed that rebamipide had good affinity with protein kinase B1 (Akt1) protein, FoxO protein, and phosphatidylinositol-3-kinase (PI3K) protein. It was confirmed that rebamipide decreased the mRNA expression of pro-inflammatory factors [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon γ (IFN-γ), interleukin-1β (IL-1β)]. The mRNA expressions of anti-inflammatory factors [interleukin-4 (IL-4), interleukin-10 (IL-10)], gastrin, and H+/K+ -ATPase were increased, and the mRNA levels of PI3K and AKT in FoxO signaling pathway were up-regulated, while FoxO levels were down-regulated. Conclusion Rebamipide may regulate the apoptosis process and inhibit inflammatory response by regulating FoxO pathway, so as to effectively relieve the gastric mucosal injury of mice with chronic atrophic gastritis.

R965

河南省医学科技攻关计划项目（201702121）