血瘀证是高血压病的常见证型，以往研究中，我们观察了高血压病血瘀证患者血清对血管内皮细胞（VECs）的损伤并初步探讨了高血压病血瘀证的形成机制，但高血压病是以血压升高为主要临床表现、与流体力学密切相关的疾病，体外静态培养的VECs并不能完全符合高血压病患者活体细胞的特性，尤其是血流机械力导致的VECs病理改变。本研究以中医整体观、恒动观为指导，在可控压力变化VECs流室系统平台上，对流场内培养的VECs加载不同强度的压力（剪切力、静水压、环张力）信号，运用流体力学、分子生物学、多元回归统计等技术，探讨VECs在不同压力加载下的"机械力因素-强度-时间-血瘀证效应"关系，并予机械力作用及活血中药干预后双向凝胶电泳筛选差异蛋白，质谱鉴定，血瘀证患者血浆验证，揭示机械力在血瘀证形成中的途径，探讨其在血瘀证形成中的机制，为以后研究奠定基础。

Blood stasis syndrome is one of the most Common syndromes of hypertension disease, in the last work, we have studied the damage of vascular endothelial cells caused by the serum of blood stasis syndrome patients with hypertension disease, and partly discussed the the influencing mechanisms in the development of blood stasis syndrome(BSS). However, hypertension is high blood pressure as the main clinical manifestations and fluid dynamics-related disease, VECs in vitro static culture and do not fully comply with the characteristics of the living cells of patients with hypertension, particularly the pathological changes caused by mechanical factor. The study is guided by the whole concept and constant dynamic concept of TCM, at the system platform of controlled pressure, VECs will bd loaded a different strength of the pressure(shearstress，pressure，strain) signal, then ,Use of fluid dynamics, molecular biology, and multivariate statistical techniques, to analysis the cell activity, morphology, ultrastructure, secretory function expression of the VECs under different pressure, and discuss the relationship Of "hemodynamic factors-strength-time-effect of blood stasis syndrome", to disprove the role of hemodynamic factors in the formation of BSS with Chinese medicine have activating blood function,to create a new cell model of blood stasis syndrome with hypertension disease, lay the foundation for future study of BSS associated with hypertension disease.

4 年来，在国家自然科学基金的资助下，本课题组对血流机械力在高血压病血瘀证形成中的机制进行研究。通过本项目的研究已发表SCI文章3篇，在投稿SCI文章1篇，已发表国内期刊5篇。以下为本研究主要发现：① 高流体静压低剪切力血流动力学条件与高血压病血瘀证状态相近。② 高血压病血瘀证血流动力学环境能诱导内皮细胞功能障碍。③ 采用iTRAQ蛋白质组学技术发现152个差异蛋白，KEGG通路富集分析发现AMPK 通路可能与高血压病血瘀证血流动力学环境诱导内皮功能障碍有关。④ 高血压病血瘀证血流动力学环境能抑制内皮细胞AMPK的活性。⑤ 高血压病血瘀证患者血清能诱导内皮功能障碍抑制AMPK的活性。⑥ 高血压血瘀证大鼠模型的主动脉组织AMPK活性下降。⑦ 丹参减少高血压病血瘀证血流动力学环境或者高血压病血瘀证患者血清诱导的内皮功能障碍，升高AMPK活性。⑧丹参抑制高血压血瘀证大鼠模型的内皮功能障碍，升高大鼠主动脉组织AMPK的活性。⑨ 丹参能升高正常培养条件下内皮细胞AMPK的活性。