基于微生物转化模型辅助指导下的赤芍单萜苷体内过程与活性物质基础研究

As one of traditional Chinese medicines, Radix Paenoniae Rubra is very rich in monoterpene glycoside compositions, which pharmacology studies show that it has a very strong pharmacological effection on the blood system, anti-inflammatory and anti-atherosclerosis. Since biotransformations of these ingrediens are often occured in the intestinal tract, the metabolic processes of monoterpene glycosides in vivo and their dynamic behaviour are not very clear, and also the real bioactive material bases of Radix Paenoniae Rubra which play the traditional efficacy role are still unclear. This study is chiefly concerned with building up a platform for natural products microbiological transformation with fungi and intestinal bacteria, finding the biological transformation metabolites of several active monoterpene glycosides as the leading compounds and exploring the pharmacokinetic process of those active monoterpene glycosides in rats by using the metabolites as reference substances. The prototype compounds and metabolites in rats plasma are determined both in qualitative and quantitative methods through modern technology. The transportation and transformation of every single active monoterpene glycoside and the whole glycoside components are examined respectively after administration, which reveals the pharmacokinetic regular of monoterpene glycosides of Radix Paenoniae Rubra and the interaction between those components. Simultaneously, the bioactive screening of these metabolites is carried out in the assay of antioxidation for injured myocardial cell in vitro. And then these studies will contribute to specifying the pharmacodynamic material bases of monoterpene glycosides and providing theory basis and new research space for Chinese traditional medicine development.

赤芍为我国传统中药，富含单萜苷类成分，药理学研究表明其在对血液系统、抗炎及抗粥样硬化等方面具有很强的药理功效。该类成分易在肠道内发生生物转化，目前赤芍单萜苷类活性物质在体内的代谢过程及其动力学行为还不十分明了，发挥赤芍传统药效的物质基础尚不明确。本课题以真菌、肠内菌等作为转化体系，建立天然产物微生物转化平台，以赤芍中几种活性单萜苷类物质为底物进行生物转化，发现以单萜苷类为先导的代谢产物，并以此微生物转化模式获得的转化产物为对照品来深入探讨赤芍中单萜苷类活性物质在大鼠体内的药代动力学过程。通过现代技术定性定量分析大鼠血浆中原型化合物及代谢产物，分别考察各活性单萜苷单体给药及总苷整体给药后各成分的转运和转化，从而揭示赤芍单萜苷的药动学规律以及各成分之间的相互作用。同时对代谢产物进行体外对心肌细胞损伤的抗氧化作用筛选，进而阐明其药效物质基础，为中药赤芍的新药发展提供理论依据和新的研究空间。

赤芍为我国传统中药，富含单萜苷类成分，该类成分在对血液系统、抗炎及抗粥样硬化等方面具有很强的药理功效。由于其易在肠道内发生生物转化，目前赤芍单萜苷类活性物质在体内的代谢过程及其动力学行为还不十分明了，发挥赤芍传统药效的物质基础尚不明确。本课题研究开展了赤芍的化学成分研究，得到了芍药苷、芍药内酯苷、氧化芍药苷、苯甲酰芍药苷、苯甲酰氧化芍药苷和芍药新苷等一系列单萜苷类化合物单体，并应用HPLC-DAD-ELSD双检测器测定不同产地赤芍中芍药苷和芍药内酯苷的含量，并进行方法学验证，为其质量标准研究建立基础。在生物转化方面，将芍药苷进行肠内菌转化，通过液质联用在MTBE萃取层中，检测到代谢物31个，鉴定了16个，其中13个为首次在肠内菌转化中发现。将芍药苷进行肝微粒体转化，通过液质联用检测发现代谢物20个，确定了6个，均为首次在该体系中发现。同时进行了葡萄糖水解酶及短乳酸菌对芍药苷的转化。在体内药物代谢研究发面，建立了脑缺血再灌注的实验动物模型，为探讨赤芍总苷在正常动物及模型动物中体内过程，开展了总苷的药代动力学研究。为探讨芍药苷大鼠体内过程，采用液质联用分别从血样中检测出代谢物9个，确定3个；尿样中代谢物23个，确定10个，其中新的代谢物1个；粪样中代谢物15个，确定6个，其中新的代谢物3个。为进一步探讨芍药苷体内过程，将芍药苷与人血清白蛋白进行了结合研究，通过荧光及分子对接的方式探究了二者的结合常数，结合位点等。另将有关赤芍单萜苷的肝微粒体转化，肠内菌转化及大鼠体内代谢研究的液质联用分析等结果进行专利申请。同时利用该基金经费开展虫草素肝微粒体的转化工作及一些挥发油的GC-MS分析研究等。