川芎冰片药对“引经、行气、活血”协同增效分子机制研究

The combination of Rhizoma Chuanxiong and Borneol is a commonly used couplet-medicine of traditional Chinese medicine in the treatment of myocardial infarction. Their compatibility includes the "guide effect", "promoting Qi", "active blood", which is a concentrated expression of the compatibility of Chinese medicine theory. To combined with modern scientific interpretation to illustrate the meaning of the compatibility of Chinese medicine theory, and improve the treatment methods of myocardial infarction is important. Our previous study found that the signaling pathways of ADR, ATP, NF-κB, PPAR were regulated by the components in Chuanxiong, which leaded to cardiovascular contraction, energy metabolism, lipid metabolism and immune regulation. The "guide effect" of Borneol is related to regulate P-gp, affect the drug concentration in target organs which could show a synergistic effect with Chuanxiong. The project is planned to through the establishment of myocardial infarction model, by system biological studies to investigate the compatibility of Chuanxiong and borneol. By bioactivity-based UPLC-Q / TOF to rapid screen the active components of Chuanxiong and borneol. Associated marks in vivo tracking of critical components and target fishing to reveal the target-pathway network, we try to clarify the synergistic mechanism of Chuanxiong and borneol on the "guide effect", "promoting Qi", "active blood" effect, and by provide a theoretical basis and experimental evidence to improve the treatment of myocardial infarction.

川芎冰片是中医治疗心肌梗死的常用药对，二者配伍包含了“引经”、“行气”、“活血”多种治则，是中医药配伍理论的集中体现。将其与现代科学相结合诠释其内涵，对于科学阐释中医配伍理论，完善现代治疗心肌梗死方案具有重要意义。我们前期研究发现川芎中存在调控ADR、ATP、NF-κB、PPAR等信号通路成分，药理作用与“心血管收缩、能量代谢、免疫调节及糖脂代谢”关系密切；引经药冰片药效成分可以调控P-gp，影响靶器官药物浓度，且与川芎药效成分具有协同作用。本项目拟通过建立心肌梗死模型，开展川芎冰片治疗心肌梗死系统生物学研究；结合活性导向的UPLC-Q/TOF快速筛选平台，辨析川芎冰片药对调控上述通路药效成分；并以化学生物学手段标记关键成分进行活体示踪及靶点垂钓，揭示其作用靶点、通路以及网络的关联；以期阐明川芎冰片药对“引经、行气、活血”协同增效分子机制，为完善现代治疗心肌梗死方案提供理论基础及实验依据。

心肌梗死是影响人类健康最常见疾病之一。中医药对于心肌梗死的治疗也积累了丰富的经验，形成了以行气活血为基础，以川芎、当归、丹参、冰片等中药为脉络的完整治疗体系。其中川芎、当归等活血化瘀中药常与冰片等引经药物配伍应用，使有效成分直达病所，提高疗效，是中医药配伍理论的集中体现。将其与现代科学相结合诠释其内涵，对于科学阐释中医理论，完善现代治疗心肌梗死方案具有重要意义。.在本项目中，我们以从系统生物学到化学生物学的研究理念为指导，首先对川芎冰片配伍治疗心肌梗死的整体药效进行评价，并应用基因组学方法对其作用机制进行系统分析，发现其对ApoE基因敲除心肌缺血模型有较好的改善作用，其机制与调节免疫、调节脂质代谢、钙拮抗等多种功效相关；同时利用前期建立的生物活性导向UPLC-Q/TOF快速筛选平台对其调控NF-κB、Ca2+、ATP、cAMP成分进行筛选，发现阿魏酸、藁本内酯、川芎嗪等调控NF-κB成分10种，阿魏酸、藁本内酯、洋川芎内酯I等钙拮抗成分5种，但未检出调节ATP、cAMP活性成分；进一步的药代动力学实验也表明，冰片可以增加藁本内酯、阿魏酸等关键药效成分在心脑等重要靶器官的分布。鉴于藁本内酯在上述两种活性中均有重要地位，我们对藁本内酯及冰片进行结构修饰，合成光敏探针用于其直接作用靶点捕获，并采用蛋白组学方法对其靶点进行鉴定，并结合多种分子生物学手段进行功能确证，发现藁本内酯可以通过直接调控SMAD3、CAMKⅡ等靶蛋白发挥调节免疫、钙拮抗作用，而冰片引经作用与直接调控RTK蛋白，促进关键药效成分吸收有关。最后采用网络药理学方法将上述结果进行关联，初步构建川芎冰片配伍“活性成分-直接靶点-通路-药效”网络。为系统诠释川芎冰片配伍“引经、行气、活血”协同增效分子机制，为完善现代治疗心肌梗死方案提供理论基础及实验依据。