基于“炭药苦味-药效物质-止血效应”相关性的蒲黄炭炮制机理研究

Changes in herbs’ taste are one of the most important aspects of the processing, because that indicates the changes of chemical constituents and medical efficacy. By literature research and experimental investigation, we found that drugs can generate bitter taste in carbonizing process. Researches about bitter taste receptors have shown that they involved in hemostatic effect. Carbonized Typhae Pollen is a commonly used herb medicine in vitro and vivo for hemostasis, with obvious bitter taste. On the basis of the existing research foundation, this study focused on “Tan Yao Zhi Xue” (means the carbonizing processed herbs have hemostatic effect), one of an important processing theory of Traditional Chinese Medicine, and proposed the hypothesis that “the bitterness could be generated during carbonizing processed; while the bitterness components can launch hemostatic mechanism, via bitter taste receptors and the bitter taste effect”. In this study, carbonized Typhae Pollen was taken as the research objects, to reveal the material basis of bitterness in carbonizing processed herbs and the mechanism of their hemostatic effect. Artificial odor (smell) bionic technology was applied to build a methodology for preliminarily screening the bitter substances in carbonizing processed herbs. Computer virtual screening technology was used to find the bitter substances which could not only bind to bitter taste receptors (TAS2Rs) but also dock with hemostatic-related receptors (eg.GPVI receptor, etc). The hemostatic mechanism of these carbonizing processed herbs could be partly explained by analyzing the docking regularity of bitter substances with the two types of receptors. The biology experiments that determine the hemostatic effect of bitter substances were applied to validate the correlation between bitter taste and hemostatic effect of carbonized Typhae Pollen. In this study, material basis of bitterness effect and hemostatic effect of carbonized Typhae Pollen could be revealed. Mechanism of “Tan Yao Zhi Xue” processing theory could also be clarified from the perspective of the correlation between “bitter taste-medicinal effective components-hemostatic effects”. This could develop a new method for the mechanism research on traditional processing of TCM and provide a demonstration for the material basis and mechanism research of efficacy of carbonizing processed herbs.

炮制对药味的影响是炮制发挥作用的重要方面。通过实验考察和文献调研发现，药物制炭后能够产生明显的苦味。而课题前期研究发现，苦味受体与止血作用存在一定联系。基于现有基础，本课题针对“炭药止血”这一炮制理论，以临床常用止血药蒲黄炭为例，提出“炭药经炮制产生苦味物质，苦味物质在作用于苦味受体产生苦味的同时，启动凝血机制发挥止血作用”的假说。课题以蒲黄炭为模型药物，采用人工味觉仿生技术建立炭药中苦味物质的感官表征方法，以发现炭药中的苦味物质；运用计算机虚拟筛选技术筛选出其中能与苦味受体TAS2Rs结合并与GPVI受体等止血相关受体作用的物质，分析苦味物质与两类受体作用的规律性；结合生物学实验，验证苦味物质的止血效应，探索炭药苦味与其止血作用的相关性。以期从“炭药苦味-药效物质-止血效应”的相关性来阐明“炭药止血”的炮制机理，开拓炮制机理研究的新思路，对系列炭药的药效物质及作用机理研究具有示范意义。

炮制在对药物功效产生影响的同时，也影响药物的药味，而药物的药味与其功效是密切相关的。药物制炭后能够产生明显的苦味，而炭药止血则是炮制学中最早的炮制理论。课题前期研究发现，苦味受体与止血作用存在一定的潜在联系。本课题提出“炭药经炮制产生苦味物质，苦味物质在作用于苦味受体产生苦味的同时，启动凝血机制发挥止血作用”的假说。课题以蒲黄炭为模型药物，采用人工味觉仿生技术建立炭药中苦味物质的感官表征方法，结合中药组分和成分分离，探索炭药中发挥苦味的主要物质；运用计算机虚拟筛选技术筛选出其中能与苦味受体结合并与止血相关受体作用的物质，分析苦味物质与两类受体作用的规律性；运用生物学实验，验证苦味物质的止血效应，探索炭药苦味与其止血作用的相关性。结果发现，蒲黄炮制后山萘酚、槲皮素、异鼠李素、柚皮素等成分含量增加，而这些物质，具有较高的苦味贡献度，并与苦味药效团结合。在与凝血酶的对接位点的虚拟筛选过程中，发现这些成分也有较好对接效果。通过血小板凝集实验验证，这些物质具有较好的止血活性。通过本课题的实验研究，对课题前期的假说进行了验证。从苦味角度，阐明了蒲黄炭止血的物质基础，对炭药的药效物质及作用机理研究，以及药物气味与功效之间关系研究奠定了基础。