基于AMPK途径探索高血压血瘀证血管内皮功能障碍及丹参防治的机制

Vascular endothelial dysfunction is the initiating factors of the process form hypertension to atherosclerosis. Blood stasis syndrome (BSS) is the common TCM syndrome of hypertension, and is high correlation with vascular endothelial dysfunction. In our preliminary experiment, we found the vascular endothelial dysfunction was present both in the cell and animal model of the BSS in hypertension, the mechanism may be related to inhibition of AMPK; Danshen (DS) inhibited the vascular endothelial dysfunction and activated AMPK both in the cell and animal model of the BSS in hypertension; 18 candidate compounds of DS is potential AMPK agonist. Thus, we put forward the hypothesis that AMPK might play an important role in BSS induced-vascular endothelial dysfunction in hypertension; DS might prevent BSS induced-vascular endothelial dysfunction through activation of AMPK; 18 candidate compounds of DS should be the chemical base of the treatment mechanism of Danshen. In present study we will determine the involvement of AMPK in the BSS induced-vascular endothelial dysfunction in hypertension by using AMPK activator or overexpression of AMPK. Next, we will determine the involvement of AMPK in the anti-vascular endothelial dysfunction actions of DS by using AMPK inhibitor and RNA interference of AMPK. Moreover, we will detect the chemical base of the treatment mechanism of Danshen by molecular simulation, enzymatic activity assay and pharmacology experiment. This study is aim to investigate the mechanism of BSS induced-vascular endothelial dysfunction in hypertension, explore the effective drugs and illuminate its mechanism and chemical base.

血管内皮功能障碍是高血压病患者发生动脉粥样硬化的始动因素，血瘀证是高血压病的常见证型，二者高度相关。本项目前期研究发现高血压病血瘀证血管内皮功能障碍可能与AMPK抑制有关，丹参能激活AMPK并抑制内皮功能障碍，二氢丹参酮I等18个丹参成分是潜在的AMPK激动剂。基于此我们提出科学假说：AMPK可能在高血压病血瘀证血管内皮功能障碍中发挥重要作用，丹参可能通过激活AMPK抑制血管内皮功能障碍，18个丹参成分可能是丹参激活AMPK防治血管内皮功能障碍的物质基础。本项目将采用激动剂或基因过表达技术激活AMPK后验证AMPK在高血压病血管内皮功能障碍中的作用；采用抑制剂或基因干扰技术抑制AMPK后验证AMPK在丹参防治血管内皮功能障碍中的作用；采用分子模拟、酶活性等技术探索丹参作用的物质基础。本项目旨在探索高血压病血瘀证血管内皮功能障碍的机制，发掘有效治疗药物并阐明其作用的物质基础。

4 年来，在国家自然科学基金的资助下，本课题组对基于AMPK途径探索高血压血瘀证血管内皮功能障碍及丹参防治的机制进行研究。通过本项目的研究已发表国内期刊中文文章1篇，SCI论文1篇，申请发明专利1项。.以下为本研究主要发现：①高血压病血瘀证患者血清能诱导内皮功能障碍和炎症反应，其机制可能与抑制 AMPK 的活性有关。②丹参能够改善高血压病血瘀证患者血清诱导的内皮功能障碍，其机制可能与激活AMPK有关。③采用计算机辅助药物筛选技术探索丹参激活 AMPK 的物质基础，发现二氢丹参酮I等18个化合物可能是丹参激活AMPK 抑制高血压病血瘀证血管内皮功能障碍的物质基础。④丹参素钠改善Ang II诱导的内皮舒张功能障碍，抑制单核-内皮黏附活动。⑤丹参素钠通过激活AMPK发挥保护内皮功能和抑制血管内皮黏附的效应⑥紫丹参甲素能够在体内外改善Ang II诱导的内皮舒张功能障碍⑦紫丹参甲素通过激活AMPK发挥保护内皮功能和抑制血管内皮黏附的效应。