目的 運(yùn)用網(wǎng)絡(luò)藥理學(xué)和實(shí)驗(yàn)探究瑞巴派特改善慢性萎縮性胃炎小鼠的作用靶點(diǎn)及通路。方法 通過(guò)PubChem中進(jìn)行檢索得到瑞巴派特的詳細(xì)信息，并分別導(dǎo)入Targetnet、SwissTarget Prediction、Super-PRED、PharmMapper數(shù)據(jù)庫(kù)進(jìn)行檢索得到與瑞巴派特作用相關(guān)的靶點(diǎn)。通過(guò)在Pharmgkb、OMIM、GeneCards、Disgnet、CTD數(shù)據(jù)庫(kù)篩選出慢性萎縮性胃炎相關(guān)的疾病靶點(diǎn)。利用韋恩圖獲取藥物與疾病的交集靶點(diǎn)，將獲得的交集靶點(diǎn)導(dǎo)入STRING數(shù)據(jù)庫(kù)，建立蛋白相互作用（PPI）網(wǎng)絡(luò)圖，借助DAVID進(jìn)行基因本體論（GO）功能、京都基因和基因組百科全書（KEGG）通路富集分析，最后對(duì)可能的核心靶點(diǎn)進(jìn)行分子對(duì)接驗(yàn)證。采用幽門螺桿菌和亞硝酸鹽建立慢性萎縮性胃炎小鼠模型，通過(guò)蘇木素–伊紅（HE）染色、免疫組化染色法、實(shí)時(shí)熒光定量（RT-qPCR）法驗(yàn)證瑞巴派特對(duì)慢性萎縮性胃炎的炎癥因子和部分關(guān)鍵靶點(diǎn)的調(diào)控作用。結(jié)果 通過(guò)數(shù)據(jù)庫(kù)篩選出相關(guān)藥物靶點(diǎn)506個(gè)，疾病靶點(diǎn)2 115個(gè)，“藥物–疾病”交集靶點(diǎn)169個(gè)，共獲得20個(gè)核心靶點(diǎn)、10類生物過(guò)程，凋亡及叉頭轉(zhuǎn)錄因子（FoxO）信號(hào)通路等20條相關(guān)通路，分子對(duì)接顯示瑞巴派特與蛋白激酶B1（Akt1）蛋白、FoxO蛋白、磷脂酰肌醇-3-激酶（PI3K）蛋白親和力良好。實(shí)驗(yàn)證實(shí)了瑞巴派特降低了促炎因子[白細(xì)胞介素-6（IL-6）、腫瘤壞死因子-α（TNF-α）、γ干擾素（IFN-γ）、白細(xì)胞介素-1β（IL-1β）]的mRNA表達(dá)，升高抑炎因子[白細(xì)胞介素-4（IL-4）、白細(xì)胞介素-10（IL-10）]、胃泌素和H⁺/K⁺-ATP酶的mRNA表達(dá)，并上調(diào)FoxO信號(hào)通路上的PI3K、AKT mRNA水平，下調(diào)FoxO水平。結(jié)論 瑞巴派特可能通過(guò)調(diào)控FoxO通路，調(diào)節(jié)凋亡過(guò)程、抑制炎性反應(yīng)，進(jìn)而有效緩解慢性萎縮性胃炎小鼠胃黏膜損傷。

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

河南省醫(yī)學(xué)科技攻關(guān)計(jì)劃項(xiàng)目（201702121）