基于体内动态效应物质变化规律的酒当归炮制机理研究

Based on the medica property theory of wine-baked radix Angelica sinensis, we previously carried out the correlation study on the compositions change of phthalides and anticoagulant activities of wine-baked radix Angelica sinensis. The results displayed that the compositions were found to exist the difference, and the anticoagulant activity was strengthened after Radix Angelica sinensis processing with wine. Traditional Chinese Medicines (TCM) were usually administered by oral, and finally the effective compounds were possible the pharmaceutical ingredients passing into the bloodstream, their metabolites and endogenous active substances. However, the effective substances and their change rules of wine-baked radix Angelica sinensis in vivo were not clear until now. Many studies suggested the medica property of TCM was transformed when the compositions changed in vivo. Therefore, we put forth a hypothesis that the compositions of wine-baked radix Angelica sinensis can be transformed or interacted in vivo due to the gastrointestinal tract absorption barrier. The absorption, transport and bioavailability of the compositions of wine-baked radix Angelica sinensis are also affected so that the clinical application of wine-baked radix Angelica sinensis is changed. The main research contents of this project include four parts. Firstly, study on the variation of the components in radix Angelica sinensis processed with wine or not; Secondly, spectrum-activity relationship of activating blood circulation components in radix Angelica sinensis processed with wine or not; Thirdly, study on the change rules of effective compositions in vivo of radix Angelica sinensis processed with wine; Finally, study on the molecular mechanism of invigoration blood in radix Angelica sinensis processed with wine. These researches will provide a scientific basis for clarifying the processing mechanism of wine-baked radix Angelica sinensis.

基于对酒当归传统药性理论的认识，我们前期开展了当归酒炙前后苯酞类成分变化与抗凝血方面的研究，发现了酒炙会引起当归中化学成分变化及抗凝血活性增强。中药多经口服给药，最终发挥药效的可能是进入血液的药物成分或其代谢产物，以及内源性活性物质。然而酒当归在体内发挥作用的效应成分及其动态变化规律尚不明确。本项目基于中药化学成分的体内过程可能导致其药性改变的现代认识，推测：酒当归的化学成分进入机体后，在胃肠道等吸收屏障的调控及其耦联作用下发生成分转化或相互作用，影响了成分在体内的吸收转运及生物利用度，从而改变酒当归的临床疗效，这可能是其活血增效的原因。故本项目拟开展以下工作：（1）当归酒炙前后化学成分/组分变化规律研究；（2）当归酒炙前后活血祛瘀的谱效关系研究；（3）当归酒炙前后效应成分的体内动态变化规律研究；（4）当归酒炙活血增效的分子机制研究。通过以上研究，以期为阐明酒当归炮制的科学内涵提供依据。

当归具有补血活血之功效，中医理论认为：当归酒炙后活血祛瘀功效增强。课题组前期研究发现：酒当归总提物体外抗凝血活性优于生当归总提物；然而酒当归效应成分变化规律、药动学差异及活血增效的分子机制尚不明确，故本项目开展了以下研究：（1）当归中化学成分及生物活性的进一步研究：从中分得4个新苯酞二聚体和3个已知苯酞二聚体，1个新的阿魏酸衍生物和5个已知酚酸类成分，它们表现出一定的COX-2抑制活性或延长TT活性。（2）当归酒炙前后不同部位活血功效的差异性研究：酒当归不同部位在降低模型鼠全血粘度、血浆粘度、血栓干湿重、血栓形成堵塞率、HTC、FIB及升高HBX，延长TT等方面优于生当归同部位，表明酒炙确实会引起生、酒当归活血功效的改变；（3）当归酒炙前后活性部位成分变化规律研究：对生、酒当归的二氯甲烷部位（ASD、WASD）定性检测结果显示，共有化学成分25个，差异性化学成分11个；以其中11个主要指标性成分定量分析发现酒炙后藁本内酯等4个苯酞类呈下降趋势；而阿魏酸等7种指标性成分呈上升趋势；对生、酒当归水部位多糖含量研究发现，2种样品中总多糖含量接近，但中性及酸性多糖含量差异显著；（4）生、酒当归中藁本内酯在不同机体状态下药动学研究：生、酒当归中藁本内酯无论在正常大鼠或模型大鼠体内，多个药动学参数存在显著性差异，综合分析发现酒当归中藁本内酯的生物利用度高于生当归，推测这可能是其活血增效的机制之一。（5）基于网络药理学和实验验证探讨酒当归活血增效的作用机制：网络药理学预测表明当归主要通过TNF、VEGF/VEGFR2、PI3K-AKT、MAPK等信号通路发挥抗炎、抑制血管生成等活性而实现活血功效；研究发现ASD、WASD是其体内外抗炎的活性部位；主要通过抑制炎症相关信号通路的激活发挥作用，相同剂量下WASD活性强于ASD，提示抗炎可能是酒当归活血增效的机制之一。本项目从药效-成分变化-体内药动学-分子机制多个角度揭示当归酒炙活血增效的炮制机理，为进一步指导合理临床用药奠定基础。