高良姜对NSAIDs引起大鼠溃疡性胃损伤的保护作用及内在机理研究

Gastric ulcer caused by non-steroidal anti-inflammatory drugs (NSAIDs) is common drug adverse reaction. Recent years, we focused on relieving NSAIDs-induced gastric damage through co-administration of Chinese medicine and NDAIDs. Our preliminary results revealed that the ethanol extract of rhizome from Alpinia officinarum Hance (AOE) could reduce ulcer index, gastric mucosa damage induced by aspirin. AOE also inhibited inflammatory factor and up-regulated the prostaglandin E2. However, the underlying protection mechanisms of AOE from NSAIDs-induced gastropathy remain largely unknown. This application aims to investigate the mechanisms of AOE and its main active components which reduce the gastric damage in murine models according to the pathological mechanisms caused by NSAIDs, including aspirin, celecoxib, naproxen and indometacin. The current application will investigate the influence of AOE and its main components on COX, iNOS, cNOS, CSE and ECE-1 using RT-PCR and Western blotting, which are mediators of gastric ulcer and inflammation. The effects of AOE and its main components on NSAIDs-stimulated gastric acid secretion in isolated rat gastric mucosal cells are planned to investigate in this project. Also, the importance of gastric hypermotility in the pathogenesis of NSAIDs-induced gastric lesions in conscious rats will be studied using a miniature balloon which is inserted into the greater curvature of the forestomach. Meanwhile, the concentration of NSAIDs and their main metabolites in various biomatrix samples (e.g., plasma, bile, urine and feces) from model rats after oral dosing will be analyzed using ESI-interfaced LC-MS/MS methods. The methods will be used to evaluate the influence of AOE and its main components on the systemic exposure to and pharmacokinetic behaviors of NSAIDs and main metabolites after co-administration to model rats. This application plans to study the roles of Alpinia officinarum Hance from a new perspective and to elucidate the underlying mechanisms. The investigation of Alpinia officinarum Hance, which is capable of reducing gastric damage caused by NSAIDs, could be useful for clinical use for the potential management of complicated adverse reactions of NSAIDs.

非甾体抗炎药（NSAIDs）引起的胃溃疡是临床常见不良反应。申请人关注中药与NSAIDs联用对其不良反应的改善作用，前期研究证实高良姜乙醇提取物（AOE）可降低阿司匹林所致小鼠胃溃疡指数，减轻黏膜损伤，抑制炎症因子释放，确切机制需要阐明。本申请聚焦于AOE对NSAIDs引起溃疡性胃损伤保护作用的机理研究。针对NSAIDs所致胃溃疡病理表现，本项目拟借助RT-PCR等分子生物学技术，评价模型组、AOE及其主要成分组COX、iNOS、cNOS、CSE和ECE-1等信号通路中关键蛋白的mRNA及蛋白表达的变化；通过胃壁细胞培养等评价对胃酸分泌的影响；通过胃部微气球植入法观察胃部过度活动的变化；利用LC-MS/MS技术研究AOE等对NSAIDs及主要代谢物系统暴露和处置过程的影响。本课题从不同角度、多个层面阐明高良姜改善NSAIDs所致大鼠溃疡性胃损伤的作用机理，为高良姜创新发展和应用提供依据。

据报道中药与非甾体抗炎药（NSAIDs）联用可减轻NSAIDs临床使用中易引起的胃溃疡。本项目拟在初试基础上，一方面进行高良姜对NSAIDs引起胃溃疡保护作用的药效及作用机制研究。另一方面从药物代谢过程，研究高良姜对NSAIDs系统暴露、组织分布及排泄过程的影响。以期探讨AOE是否可以改善NSAIDs在临床使用中所引起的溃疡性胃损伤。. 针对NSAIDs所致胃溃疡病理表现，评价了常见非甾体药物塞来昔布、阿司匹林、萘普生和吲哚美辛致大鼠胃损伤造模效果，确定采用吲哚美辛进行造模。借助病理学检验以及western blot、RT-PCR等分子生物学技术，评价模型组、AOE及其代表性单体化合物高良姜素对吲哚美辛作用下胃粘膜的保护效果；研究COX、iNOS、cNOS、CSE和ECE-1等信号通路中关键蛋白的mRNA及蛋白表达的变化；通过胃壁细胞培养实验等评价对胃粘膜屏障的影响；利用LC-MS/MS技术研究AOE等对NSAIDs及主要代谢物系统暴露和处置过程的影响。结果表明：高良姜能够降低炎症因子水平、促进VEGF表达、维护胃粘膜血流稳定，其机制与提升COX-1酶活性、激活NO与H2S合成通路关键酶、抑制H+-K+-ATP酶活性、降低细胞膜通透性、抑制凋亡蛋白caspases 3 和caspases 9蛋白的活化、降低NSAIDs诱导的细胞凋亡水平等有关。同时，高良姜提取物可减少了吲哚美辛的系统暴露、加快消除过程，并促进吲哚美辛及其代谢物在大鼠胆汁和粪中的排泄。. 本项目从药效及作用机制、药物代谢过程两方面充分证明高良姜可以改善NSAIDs产生的胃损伤作用，为今后临床上中西药联合使用奠定科学基础，在给患者使用NSAIDs治疗炎症的同时服用高良姜汤剂以减轻不良反应；同时，高良姜为药食同源品种，可以开发以高良姜为主针对胃损伤的食品，减轻病人服用NSAIDs的不良反应，为高良姜创新发展和应用提供依据。