新疆一枝蒿抗ILI作用的药效物质基础与作用机制研究

Uygur medicine is one of the distinct traditional medicines, possessing the unique theory system, curative effect and significant characteristics. The representative Uygur medicine Artemisia rupestris L. (AR) with antivirus, liver preservation and antienzyme, anti-inflammatory, anti-allergy, antioxidation, regulation of immunity effects, has been used mainly for influenza and hepatitis. Followed the guidance of the traditional medicine theory and understanding of the whole concept of drug efficiency/efficacy for its pharmacology, based on exact curative effect and clinical application of AR, combining the data analysis and results of the previous work accumulation, we put put forward the scientific hypothesis in this project. It will be established an animal model of Con A induced immune liver injury to probe into the active material basis and molecular mechanism with integrate methodology mode of pull together interdisciplinary research methods and techniques such as UPLC-MS analysis, holistic animal efficacy evaluation, molecular biology and others. The focus of the research involve contents of active component characteristics and molecular mechanism, effects of endogenous metabolites and potential biomarker discovery, based on a systemly biological association mode of “constituents/ transitional components and their pharmacodynamics→efficacy/ molecular mechanisms→metabolomics difference / potential markers recognition” simultaneously on three dimensions. The results of research contents would interpret the against immunological liver injury effect of Artemisia rupestris on in vivo level for its pharmacodynamic substance and its molecular and metabolic mechanism, provide scientific basis of medicine theory and clinical application. The implementation of the project will expand the scope of clinical application of AR, further explore the precious medicinal resources, enhance the study of therapeutic material, and for the enhancement of innovative research of drug discovery from ethno medicine also has an important inspiration and reference.

维吾尔医药理论自成体系，药性理论独特，药物疗效与特色鲜明。本项目遵循维医药学理论和药性/药效整体观认识的指导，以维医药学中疗效确切、应用广泛的新疆一枝蒿为研究对象，结合前期已有工作基础，提出科学假说，构建ConA诱导的免疫性肝损伤实验动物模型，整合UPLC-MS分析方法、整体动物药效评价、分子生物学方法等多学科交叉研究方法与技术，从“组分/移行成分-药效/分子机制-代谢组学/潜在标志物”三个维度系统生物学关联模式，开展新疆一枝蒿抗ILI作用活性组分的体内吸收分布及代谢规律、药效作用特点与分子机制、对内源性代谢物的影响规律及潜在生物标志物发现等研究，科学阐释其ILI作用的体内药效物质及其分子与代谢层面作用机制等实质内涵。本项目预期结果为维医学药性/药效理论认识及新疆一枝蒿合理应用提供科学依据，也探索将整体思维、系统方法论与现代科学的还原性方法相结合开展民族药应用基础研究的思路与方法。

本项目在维医药学理论指导下，以疗效确切、应用广泛的典型维药新疆一枝蒿（ARL，Artemisia rupestris L.）为研究对象，应用ConA诱导的免疫性肝损伤实验动物模型，采用了UPLC-MS分析方法、整体动物药效评价、分子生物学方法等研究方法，分别从组分分析、药效与机制、体内代谢三个维度层面，探究了ARL组分的体内吸收分布及代谢规律、抗肝损伤药效作用特点与分子机制、对内源性代谢物的影响规律及潜在生物标志物发现等内容。构建了新疆一枝蒿提取物制备和质量控制方法；应用HPLC-IT-TOF-MS从ARL化学成分组中鉴定124种成分，应用shotgun和DI-MS/MSALL技术鉴定其特征化学成分组中44种成分，应用LC-QExactive-MS技术准确检识主要成分类型的体内代谢产物及其过程，分别从实验动物血浆、尿液和粪便样品中鉴定了145个代谢物，解析其体内的主要代谢途径为磺酸化、糖醛酸化、甲基化、羟基化、氢化和脱糖基化等代谢途径；应用C57BL/6J小鼠进行了ARL抗免疫性肝损伤整体动物药效评价，ALT、AST和SOD、MDA、MPO、NO和GSH-PX等血清酶学指标和病理组织切片分析结果表明ARL对实验性免疫性肝损伤小鼠具有良好的保护作用；应用UPLC-MS/MS技术分析鉴定了ARL干预免疫性肝损伤小鼠的内源性代谢物，进行了正、负离子模式下非靶标代谢组学轮廓分析，识别其中32个潜在生物标志物，解析其主要通过纠正卟啉和叶绿素代谢通路、花生四烯酸代谢通路以及甘油磷脂代谢通路的代谢机制。本项目研究结果科学阐释了新疆一枝蒿抗免疫性肝损伤作用的体内药效物质及其分子与代谢层面作用机制等实质内涵，为维药新疆一枝蒿的临床合理应用和资源开发提供科学依据。