基于代谢组学及热分析技术的地黄炭止血“存性”与制炭工艺相关性研究

About the terms of processing traditional Chinese material media such as "firing strength", "duration and degree of heating", "nature of the drug partly preserved after carbonization", It is still mostly based on experience judgement, and lack of objective, numerical standards for the description and determination,to a great extent, restricts the development of this discipline. The traditional research idea and method chiefly focus on the variation about chemicals in vitro of processed traditional Chinese material media, it can not fully reveal the integrity and complexity of the efficacy changes before and after processed. Based on the breakthrough point that the effects of partly preservation of the nature of Dried Rehmanniae Radix Carbonisata is different from Rehmanniae Radix Praeparata Carbonisata,we put forward the working hypothesis that the effects of hemostasis ＆partly preservation of the nature of Rehmanniae Radix Carbonisata depend on its material baisis concluding transformation products in vitro & metabolites in vivo. The quantitative and quanlitative change of the material baisis to a extend mainly depend on the charcoal processing crafts .Drug metabonomics technology combined with thermal analysis technology will be used to systematically study the change progress of effective substance in the process of Charcoal Processing (Carbonization by Stir-Heating or by Calcining) of Rehmanniae Radix, to quantify the process parameters; and to analyze the dynamic process and metabolites in vivo about Rehmanniae Radix and Rehmanniae Radix Carbonisata, which will further illustrate the correlation between the charcoal processing crafts ( carbonization by stir-heating or by calcining ) and the effects of hemostasis ＆partly preservation of nature of Rehmanniae Radix, and establish a new research model of processing crafts and processing mechanism of TCM based on the technology of thermal analysis and metabonomics.

中药炮制过程中有关“火力”、“火候”、“存性”等术语的表述与判定大多依然停留在经验判断基础上，缺乏客观化、数据化的标准，在很大程度上制约着本学科的发展；对中药饮片炮制过程中体外化学物质变化规律进行研究的思路与方法，不能完全反映中药饮片炮制前后功效变化的整体性与复杂性。本项目以生地炭“存性”差异为切入点，提出“地黄炭止血'存性'的物质基础，不仅包括体外转化产物，还包括体内代谢产物；而这些物质基础的量变与质变一定程度上主要取决于制炭工艺”的工作假说。通过采用代谢组学技术，结合热分析技术，对生地黄制炭（炒炭、煅炭）过程中药效物质变化过程进行系统研究，并对工艺参数进行量化表征；对生地黄炭在体内动态变化过程和代谢产物变化进行比较分析，进而阐明生地黄制炭（炒炭、煅炭）工艺与其止血“存性”之间的相关性，构建基于热分析-代谢组学技术的中药炮制工艺与炮制机制研究的新模式。

中药炮制过程中有关"火力"、"火候"、"存性"等术语的表述与判定大多依然停留在经验判断基础上，缺乏客观化、数据化的标准，在很大程度上制约着中药的发展；对中药饮片炮制过程中体外物质变化规律进行研究的思路与方法，也不能完全反映中药饮片炮制前后功效变换的整体性与复杂性。.本实验以生地炭“存性”差异为切入点，提出“地黄炭止血‘存性’的物质基础，不仅包括体外转化物，还包括体内代谢物；而这些物质基础的量变与质变一定程度上主要取决于制炭工艺”的工作假说。通过采用代谢组学技术，结合热分析技术，对生地黄（炒炭、煅炭）的工艺参数进行量化表征；对生地炭在体内动态变化过程和代谢产物变化进行比较分析，进而阐明生地黄制炭（炒炭、煅炭）工艺与其止血“存性”之间的相关性。结果发现，生地黄及其炒炭品可显著降低大鼠肛温，降低全血高、中、低切黏度和血浆黏度，缩短 APTT 和 TT，降低RBC， Hb， HCT， FIB含量，减少肺、胃组织出血性病理损伤。生地黄煅炭品仅可缩短 APTT，TT，对其他指标未有显著改变。提示生地黄炒炭前后均具有“凉血止血”作用，生地黄煅炭仅保留“止血”作用，而无显著“凉血”作用。在体外化学成分研究中发现，生地黄经过炭制之后，有效成分的含量发生了显著的变化，且两种炭制方法之间也具有一定的差异性。建立了地黄五种成分的HPLC方法和指纹图谱。血清药物化学的研究中发现性别对地黄及其制炭品入血药效成分没有显著影响。两种炭制法之间入血成分类似，但是具体含量有差别。另外生地黄制炭前后，梓醇以原形进入到血液中。热分析结果发现216.3℃为地黄炒炭最佳温度，梓醇的最大热失重速率峰峰温274.3℃为炒炭温度上限。生地黄煅炭的最佳温度应控制在果糖、梓醇热解之前，为 5-HMF 热解最大热失重速率峰时的温度，即194.2℃，梓醇的最大热失重速率峰峰温276.8℃为煅炭温度上限。本实验为生地黄及其炭制品的质量标准控制提供了方法，构建了基于热分析-代谢组学技术的中药炮制工艺与炮制机制研究的新模式。