温中药山橿干预消化性胃溃疡的活性成分及其体内代谢机理研究

Peptic Gastric Ulcer (PGU) is a common and frequently-occurring disease. Clinical epidemiology estimates the disease rate of peptic gastric ulcer and its related diseases as high as 10%, especially in the past ten years, the disease rate of peptic gastric ulcer continued to increase. Triple therapy was often used in western medicine, but there are limitations and the adverse reactions, so the scholars at home and abroad turned attention to traditional chinese medicine. The folk medicine Lindera reflexa Hemsl. has a long history in clinical practice for treating chronic gastritis and gastric ulcer. It’s the main raw material for the production of "Weitengning tablet", which has significant curative effect for gastric ulcer disease. However, the active constituents and metabolism mechanism of L. reflexa is not clear. The project intends to adopt the LC-MS technology for the rapid qualitative and quantitative characterization of chemical compositions of L. reflexa; A quality evaluation strategy for active constituents by component knock-out/knock-in strategy is set up preliminary. The active constituents were obtained by screening three aspects of anti-inflammatory, oxidative damage protection and bacterial inhibition. PK-PD analysis was used to reveal the effective components by in vivo pharmacokinetics process and pharmacodynamic action. The aim of the research was to find out the active constituents and elucidate the metabolism mechanism of L. reflexa for the treatment of peptic gastric ulcer to provide the scientific basis for the clinical application of L. reflexa.

消化性胃溃疡是临床常见病和多发病。流行病学估计消化性胃溃疡及其相关疾病发病率高达10%，尤其近十年来，消化性胃溃疡的发病率持续增加。西医常采用三联治疗方案进行治疗，但存在局限性和不良反应，所以国内外医学界越来越把注意力转向中医药。山橿在河南大别山区用于治疗胃溃疡、胃炎历史悠久，以山橿为主要原料生产的“胃疼宁片”疗效显著。然而目前对于山橿发挥疗效的物质基础及体内代谢机理尚不明确。本项目拟采用液质联用、气质联用技术，对山橿中的化学成分群进行快速定性定量表征；并建立一种化学与药效学相结合的“化学成分敲出/敲入-谱效集成表征”策略，从抗炎、氧化损伤保护及幽门螺旋杆菌抑制三个方面筛选山橿有效成分（群）；采用PK-PD综合分析技术通过其在体内的药代动力学过程和药效进行活性及作用机制验证。从而阐明山橿治疗消化性胃溃疡的物质基础和作用机理，为山橿药材质量标准的制定及治疗消化性胃溃疡药物的发现提供科学依据。

消化性胃溃疡是临床常见病和多发病。西医常采用三联治疗方案进行治疗，但存在局限性和不良反应，所以国内外医学界越来越把注意力转向中医药。山橿在河南大别山区用于治疗胃溃疡、胃炎历史悠久，以山橿为主要原料生产的“胃疼宁片”疗效显著。然而目前对于山橿发挥疗效的物质基础及体内代谢机理尚不明确。. 本项目采用LC-MS技术从山橿乙酸乙酯部位中鉴定出了42个化学成分；采用GC-MS技术从山橿挥发油中鉴定出了20个主要的挥发性成分；建立了山橿乙酸乙酯部位中12 个色谱峰所代表的化学成分的高效液相色谱含量测定方法，明确了12个成分在山橿乙酸乙酯部位中的含量，顺利完成了山橿有效部位化学成分群的快速定性定量表征。. 采用“化学敲出/敲入-谱效集成表征”的方法，制备出了山橿乙酸乙酯部位的1-12号特征峰、1-12号阴性、全阴性样品；根据消化性胃溃疡发病机理，同时以脂多糖诱导小鼠巨噬细胞为炎症模型，脂多糖诱导小鼠巨噬细胞为氧化损伤模型及抑制幽门螺旋杆菌试验进行山橿乙酸乙酯部位化学成分群活性检测，最终确定银松素-3,5-二-O-β-D-葡萄糖苷、银松素、3-甲氧基-5-羟基-反式-二苯乙烯、3,5-二甲氧基-反式-二苯乙烯、reflexanbene C、reflexanbene I、reflexanbene B、reflexanbene A8个二苯乙烯类化合物按照其在山橿乙酸乙酯部位所占的比例进行配伍，作为山橿干预消化性胃溃疡的活性成分群。. 建立了柱色谱层析、制备液相相结合的方法导向分离山橿活性成分群。通过无水乙醇、吲哚美辛和幽门结扎诱导的胃溃疡模型对筛选出的山橿活性成分群进行体内活性验证，表明所筛选出的山橿活性成分群是山橿发挥抗消化性胃溃疡作用的药效物质基础。. 建立了高效灵敏的UPLC-MS方法检测大鼠血浆、各组织及排泄物中山橿活性成分群的含量；采用PK-PD综合分析技术探明了山橿活性成分群的体内药代动力学特征，MDA、PGE2等药效学指标的动态变化过程及二者之间的相互关系。从而阐明了山橿治疗消化性胃溃疡的物质基础和作用机理，为山橿药材质量标准的制定及治疗消化性胃溃疡药物的发现提供科学依据。. 该项目发表论文6篇，其中SCI 4篇、中文核心2篇；申请发明专利4项；培养研究生5人。