基于网络药理学及实验验证探讨达原饮治疗幽门螺杆菌阳性口疮作用机制

Exploration on the Mechanism of Dayuan Decoction in Treating Hp Positive Oral Ulcer Based on Network Pharmacology and Experimental Validation

目的基于网络药理学和实验验证探究达原饮治疗幽门螺杆菌(Hp)阳性口疮的作用机制。方法利用TCMSP数据库收集达原饮的活性成分及其作用靶点,通过GeneCards数据库获取Hp阳性口疮的相关靶点,利用Venny2.1.0在线平台对药物活性成分靶点与疾病靶点取交集,使用STRING11.0数据库构建蛋白相互作用网络,并运用Cytoscape3.7.1软件筛选核心靶点,利用DAVID数据库对交集靶点进行GO和KEGG通路富集分析。采用标准Hp菌液灌胃联合冰醋酸灼烧口腔黏膜构建Hp阳性口疮大鼠模型并予达原饮干预,观察模型大鼠口腔、胃黏膜组织形态,检测核心靶点在大鼠血清及口腔、胃黏膜组织中的表达。结果经数据库检索和筛选得到达原饮治疗Hp阳性口疮靶点165个,主要活性成分为槲皮素、山柰酚、5,8,2'-三羟基-7-甲氧基黄酮等,核心靶点有AKT1、IL6、TP53、TNF等。GO和KEGG通路富集分析提示达原饮治疗Hp阳性口疮主要集中于炎症反应相关通路。动物实验显示,与空白组比较,模型组大鼠口腔和胃黏膜组织结构紊乱,炎性细胞浸润明显,血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量升高(P<0.05),口腔、胃黏膜组织Toll样受体4(TLR4)、NLRP3含量升高(P<0.05),口腔黏膜组织TLR4、核因子-κB(NF-κB)蛋白表达升高(P<0.05);与模型组比较,达原饮各剂量组大鼠口腔和胃黏膜组织病理形态明显改善,血清IL-6、TNF-α及口腔、胃黏膜组织TLR4、NLRP3含量降低,口腔黏膜组织TLR4、NF-κB蛋白表达降低(P<0.05)。结论达原饮可能通过下调TLR4、NF-κB水平调节Hp阳性口疮的炎症发展,改善其症状。

Objective To explore the mechanism of Dayuan Decoction in treating Helicobacter pylori(Hp)positive oral ulcer based on network pharmacology and experimental validation.Methods Active components of Dayuan Decoction and its targets were collected using the TCMSP database;relevant targets for Hp positive oral ulcer were screened through the GeneCards database;the Venny 2.1.0 online platform was used to screen the intersection targets of drug active components and diseases;a protein-protein interaction network was built using STRING 11.0 database and core key targets were screened using Cytoscape 3.7.1 software;GO and KEGG enrichment analysis was conducted on intersection targets using the DAVID database.Oral administration of standard Hp bacterial solution and burning of oral mucosa with glacial acetic acid were used to construct an animal model of Hp positive oral ulcer,and the model rats were intervented with Dayuan Decoction by gavage.The morphology of oral and gastric mucosal tissues was observed,and the expression of core targets in oral and gastric mucosal tissues were detected.Results A total of 165 targets were obtained for the treatment of Hp positive oral ulcer with Dayuan Decoction.The main active components were quercetin,kaempferol,5,8,2'-trihydroxy-7-methoxyflavone,etc.,the core targets were AKT1,IL6,TP53,TNF,etc.GO and KEGG enrichment analysis suggested that Dayuan Decoction in the treatment of Hp positive oral ulcer was mainly related to the inflammatory pathway.Animal experiments showed that compared with the blank group,the structure of the oral and gastric tissues in the model group rats were significantly disrupted,with significant infiltration of inflammatory cells;the contents of IL-6 and TNF-αin the serum significantly increased(P<0.05),the contents of TLR4 and NLRP3 in oral and gastric mucosal tissues significantly increased(P<0.05),and the protein expression of TLR4 and NF-κB in oral mucosal tissues significantly increased(P<0.05).Compared with the model group,the histopathological manifestations of rats in all doseage of Dayuan Decoction groups significantly improved,the contents of IL-6 and TNF-αin the serum,and the contents of TLR4 and NLRP3 in the oral and gastric mucosal tissues significantly decreased,the protein expression of TLR4 and NF-κB in the oral mucosal tissue significantly decreased(P<0.05).Conclusion Dayuan Decoction is possible to regulate the inflammatory development of Hp positive oral ulcer and improve their symptoms by downregulating TLR4 and NF-κB levels.