基于吸收代谢的玄参加工炮制机制研究

The main bioactive constituents of Scrophulariae Radix, Harpagoside, decreased in the process of conventional processing method. According to the theory of Traditional Chinese Medicine, however, the majority of Chinese materia medica in the clinical usage must be processed. It's difficulty to illustrate the mechanism of the processing according to current standards of Scrophulariae Radix, so our research hypothesis of this project is produced: The more absorption of bioactive constituents in Chinese materia medica will be enhanced the efficacy of clinical medicine after processing..The constituents of Chinese materia medica are complicated. After absorbing into the body, through a series of biological transformation processes the medical efficacy is ingredients group maybe consist of prototype components, metabolites, or both, and the endogenous compounds, which is the effect made by a variety of constituents. .In this project, the commonly used Chinese materia medica -Scrophulariae Radix will be measured. We will use quality control method consist of the quantitative determination of multi-index constituents and the fingerprint chromatogram by UPLC-MS/MS-MRM, based on the classical quality control methods. We will compare chemical composition information before and after processing and then compare the information of absorption and metabolism in vitro and in vivo. We can get ingredients group and as a standard to assess the quality of different processed Scrophulariae Radix through interrelated analysis and calculation, furthermore, to illuminate the processing mechanism of Scrophulariae Radix from the point of view such as the absorption and metabolism. At the same time the research will provide scientific references for the rational processing technology and quality standards of Scrophulariae Radix in future.

玄参经加工炮制后主要活性成分哈巴俄苷含量降低，但是历代临床均以传统加工炮制品药用，现行的质量标准难以解释玄参的传统加工炮制机制，这就形成了本项目的研究假说：中药经加工炮制，可提高活性成分的吸收，从而达到增强药效的目的。.中药化学成分复杂，进入机体后通过一系列的生物转化过程，在体内发挥药效的可能是原型成分、代谢产物、或二者与机体内源性物共同组成的物质群，是多种成分共同作用的结果。本研究选择常用中药玄参，在经典质量评价方法的基础上，采用多指标成分定量结合指纹图谱的UPLC-MS/MS-MRM质量评价方法，将玄参加工炮制前后化学成分信息与其体内外吸收、代谢信息相结合，经分析、计算等相关处理，提取与吸收代谢相关的成分群，并以此为指标评价玄参不同加工炮制品的质量，从活性成分吸收与代谢的角度阐明玄参的加工炮制机制。同时为制订合理的玄参加工炮制工艺和质量标准提供科学依据。

玄参的传统加工炮制方法为“发汗”法，玄参经传统加工炮制后，药材颜色变黑，主要活性成分哈巴俄苷含量降低，但是中医临床均以传统加工炮制品药用，用现行的质量标准难以解释玄参“黑色质优”的传统观念和加工炮制机制。本研究选择传统加工方法对其含量影响显著的主要活性成分及传统加工玄参、切片烘干玄参和阴干玄参，研究大鼠体内的吸收与代谢和分布，从活性成分吸收与代谢的角度阐明玄参传统加工炮制机制。. 建立了玄参中哈巴俄苷、哈巴苷、桃叶珊瑚苷、6-O-甲基梓醇、斩龙剑苷A、肉桂酸等主要成分的含量测定方法和玄参药材HPLC指纹图谱，研究了不同加工方法对玄参中主要成分的含量影响。采用HPLC-MS技术对玄参化学成分进行分析，共指认玄参化学成分64个，玄参中首次报道的化合物19个；新化合物2个。采用UPLC /Q-TOF-MS技术对烘干玄参和传统加工玄参进行分析比较，结果与烘干玄参相比，传统加工玄参中共有26种成分含量变化显著（含量升高的7种，含量降低的19种）。. 采用HPLC-MS分析技术对传统加工炮制玄参、烘干玄参、哈巴俄苷、哈巴苷等进行大鼠体内吸收、代谢和分布研究，鉴定了体内可吸收成分和代谢产物，推测了各类型成分的主要代谢途径。采用UPLC/Q-TOF-MS技术、Progenesis QI代谢组学生物信息处理软件等进行数据分析，检测到烘干玄参与传统玄参给药大鼠体内差异性成分218种，血液中86种，尿液99种，粪便64种。与烘干玄参相比，传统加工玄参给药组体内原形成分和主要代谢产物含量较低，内源性成分含量较高，推测传统加工玄参给药后会对大鼠的体内自身代谢产生较大影响，通过影响体内内源性成分变化而发挥药效。. 对传统加工玄参和阴干玄参进行药理实验，发现传统加工玄参的抗菌活性优于阴干玄参，两者有显著性差异；传统加工玄参和切片烘干玄参均能显著抑制二甲苯所致的小鼠耳肿胀和角叉菜胶致小鼠足跖肿胀，两者无显著性差异，发现相同剂量条件下烘干玄参具有致泻作用，因此推测玄参经传统加工炮制后并未影响其抗炎活性，还具有一定的减毒作用（防止腹泻）。