基于生物力药理学的丹参药效机理研究

Salviae Miltiorrhizae has been used as a traditional activating blood circulation medicine for thousands of years, and its main components have been validated by modern medicine. Here we use salviae miltiorrhizae and its main components as tool medicines for activating blood circulation study. The study sought to test the hypotheses that "biomechanopharmacological mechanism is the important basis of pharmacodynamics of salviae miltiorrhizae and its main components".Factorial design with two factors (drug dosage and shear stress as two independent variables) and multi-levels is used to test the hypotheses.Bioflux1000 live cell assays with controlled shear flow system is employed to study the endothelial bioresponses induced by combination of shear stress and salviae miltiorrhize or its main components dynamically.Blood stasis syndrome animal model is established, and quantitative physical exercise is used to modulate blood shear stress. Effects of salviae miltiorrhizae or its main components combined with shear stress on vascular endothelium function are studied to explore their activating blood circulation mechanism through biomechanopharmacological methods. A comprehensive analysis of relationships between shear stress, vascular endothelial responses, drug dose and efficacy are performed by means of nonlinear statistics.This study is launched to explore new way for caridiovascular disease prevention and treatment by means of biomechanopharmacology. It will deepen the understanding of TCM theory of activating blood circulation and promote the development of integrated traditional and western medicine in the near future.

本项目选临床疗效确切且研究基础好的中药丹参及其主要成分为活血化瘀代表药物，提出的科学假设是"生物力药理学机理是丹参及其主成分的重要药效基础"。利用Bioflux1000控剪应力微流培养系统及活细胞显微缩时摄影工作站，采用双因素（药物剂量和血流剪应力作为自变量）多水平析因设计，实时、动态地研究丹参及其成分与剪应力共同参与下的内皮细胞生物学效应。并采用血瘀证动物模型，以定量运动调整血流剪应力水平，研究丹参与血流剪应力对于血管内皮功能的联合效应，揭示丹参活血化瘀作用的生物力药理学机理。采用多元非线性统计方法，综合分析血流剪应力、血管内皮细胞功能、药物剂量和药效之间关联与规律。项目从生物力药理学角度探索防治心脑血管疾病新途径，有助于深化对活血化瘀的认识，促进中西医结合研究的发展。

血流力学因素与生俱来并且无处不在，血流剪应力具有多靶向药物样作用。我们课题组基于“气血流通，百病自已”的传统中医理论，对活血化瘀中药丹参、三七的生物力药理学研究证明：一方面，生物力学因素影响和改变了经典的药理学剂量-效应关系；另一方面，在流动力学条件下，可以提高活血化瘀药效评价的敏感性。.本课题研究发现在给予中等临床剂量丹参提取物的前提下，联合定量游泳锻炼在调整血管内皮功能，在改善血液“浓、黏、凝、聚”等方面，特别是降低血小板聚集性、血浆纤维蛋白原活性，抗氧化等方面具有协同作用，从而提高老年血瘀大鼠微循环血流灌注量，改善微循环障碍。研究发现生理流动剪应力有利于丹酚酸B发挥抗血小板聚集效应，剪应力可以促进丹酚酸B诱导内皮细胞分泌PGI2。在力学环境下，筋骨草提取物乙酰哈巴苷抗炎、抗白细胞粘附、抑制内皮细胞损伤修复的效应，较静止条件的药效学更为显著。.血小板本身就是一个力学感受器，血流剪应力在抗血小板药理学研究方面具有关键作用。本项目采用“量反应平行线”法，建立了三七总皂苷体外抑制血小板聚集的生物活性测定法用于补充其质量控制方法。研究结果发现三七总皂苷对凝血酶诱导的血小板聚集具有显著抑制作用，并且呈现出剂量依赖关系，效价测定稳定并具有可重复性。利用BioFlux 1000控剪应力微流培养系统模拟体内流动条件，建立TNF-α诱导的血管内皮细胞损伤模型，研究显示生理和病理流动条件下三七均可抑制血小板粘附，抑制率分别为15.0%和34.1%。在不同力学环境下，血栓通胶囊对血管内皮细胞炎症蛋白及粘附蛋白表达有不同的影响。在病理低剪应力或静态条件下，药物能够明显抑制血管内皮细胞VCAM-1的表达和TXB2释放。而在正常生理流动下，药物对VCAM-1表达无明显影响，但是能够显著抑制内皮细胞膜蛋白VE-cadherin、细胞间连接蛋白Cx43的表达，通过阻断炎症反应导致的血管内皮细胞间粘附连接结构破坏，降低细胞通透性，保护内皮细胞单层的完整性。力学环境有助于发现抗血小板聚集的中药及其组合物。