基于多维谱效和入血成分追踪研究山蜡梅叶的物质基础

Chimonanthus nitens is the genuine medicinal materials of Jiangxi, is Chinese endemic piant. Dry leaves of Chimonanthus nitens is considered as tea for treating common cold, influenza by the indigenous people, is Jiangxi's main products. At present, the research on material basis and quality control of Chimonanthus nitens is little. Under the guidance of Chinese medicine theory, this topic using different extract solvents to prepare various samples, then conducting antitussive, anti-inflammatory, antimicrobial and antipyretic pharmacological tests. Based on the pharmacological results, a variety of instrumental analysis methods and preparation conditions was introduced to set up a plurality of fingerprints, which can reflect the traditional Chinese medicine chemical characteristics, association analysis was performed on Multi-dimensional spectral data and pharmacological results by Chemical statistics and computer technology in order to determine the effective components group, the chemical structure of the effective components group was separated、identified and characterized by LC-MS and column chromatography. At the same time, the monomer of clear structure was gavaged in rats, to study the metabolic mechanism of these monomer component, analysis of chemical composition and identification of its prototype components and metabolites were based on oral medicine extract in rat plasma. Through the research of the combination of in vitro and in vivo, pharmacodynamic material can be verified and supplied. This is very important for the pharmacodynamic material basis and quality control of Chimonanthus nitens, laying the foundation for the further research on its mechanism and the further development and utilization.

山蜡梅叶是江西的道地药材，是中国特有的植物，当地民间习惯用其叶泡茶治疗感冒和流行性感冒，也是江西主特产品。目前对其物质基础和质量控制研究很少。本课题在中医理论的指导下，制备山蜡梅叶不同溶剂提取物样品，分别进行镇咳、抗炎、抑菌和解热药理试验，在药理结果的基础上，采用多种仪器分析方法和条件制备获得能反映中药化学特性的多张指纹图谱，通过化学统计学和计算机技术将多维谱图数据和药理结果进行相关联分析，确定有效成分群，并采用液质联用和柱层析色谱对有效成分群的化学结构进行分离鉴定和表征。同时将这些结构明确的单体化合物分别灌胃给药大鼠，研究它们在大鼠体内的代谢规律，在此基础上对口服山蜡梅药材提取物后大鼠血中的化学成分进行解析并鉴定其原型成分及代谢产物。通过由“体外到体内，从单体到整体”的研究，可以对药效物质相互验证和补充，真正阐明其物质基础；也为深入研究其作用机制和进一步开发利用奠定基础。

山蜡梅是江西的道地药材，是中国特有的植物，民间习惯用其叶泡茶治疗感冒和流行性感冒。本课题在中医理论的指导下，对山蜡梅叶的药效物质基础进行了研究，具体研究内容和结果如下：（1）从山蜡梅叶75%乙醇提取液中首次分离鉴定了3个黄酮苷化合物，分别为山奈酚-3-o-β-D-半乳糖-(6-1)-α-L-鼠李糖苷，山奈酚-3-o-芸香糖苷和紫云英苷；（2）采用正交试验，优化提取和制备工艺，制备山蜡梅配方颗粒，并对颗粒剂中芦丁、东莨菪内酯、滨蒿内酯、异嗪皮啶进行了定性鉴别，对芦丁进行了含量测定；（3）采用高效薄层色谱二次展开技术对山蜡梅叶中3个萜类物质（芳樟醇、桉油精和β-石竹烯）和其他5个成分进行了定性鉴别和定量测定；（4）采用HPLC-PAD进行了化学成分色谱分离并对8种成分进行了含量测定，并建立了24批次山蜡梅叶的指纹图谱，运用SIMCA 13.0软件进行主成分分析，不同地理来源的山蜡梅叶有差异性。（5）用不同极性溶剂提取山蜡梅叶，结果表明山蜡梅叶不仅具有抑菌、解热、抗炎的作用，乙酸乙酯部位有较强的镇咳作用；用二甲麦角新碱、纳洛酮和格列本脲探讨作用机制，结果显示可能是通过中枢作用起到镇咳作用。（6）通过灌胃给药大鼠，采取UPLC-QTOF-MS/MS定性鉴别了大鼠血清中5种化学成分，分别为异嗪皮啶、东莨菪苷、东莨菪内酯、山奈酚和槲皮素，将体外有效成分群和体内代谢物比较，初步推测，该5种成分可能是山蜡梅叶的药效物质基础。上述研究成果对进一步研究山蜡梅叶防治感冒和流行性感冒的分子机制及控制山蜡梅叶的质量都有重要的实际意义。.

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