红花-乳香药对"相须""相使"药效物质及其调控NO-cGMP、 NF-kB与Nrf2的协同作用机制研究

Scientific connotation of relations among TCMs was still unclear. The diseases caused by "Blood Stasis Syndrome" were the leading diseases worldwide. Safflower-Frankincense who was the core components in hundreds of Chinese medicinal formulaes was widely used for treatment and prevention of these diseases（The sale of "Bu Chang Nao Xin Tong" was up to 2 billion). However,active components and pharmacological mechanisms of Safflower-Frankincense were unkonwn. We identified that hydroxy safflower yellow A (HSYA) and boswellic acid (BA) were two main active components of Safflower and Frankincense by using (Differential efficacy of serum chromatographic analysis，DESCA). In preliminary experiments, we found that both HSYA and BA modulated blood circulation, anti-inflammation and anti-oxidation through regulating NO-cGMP, NF-kB and Nrf2 mediated pathways. Thereby, we suppose that Safflower-Frankincense show pharmacological effects through synergetic regulation of these molecular pathways. Here, we are sight to identify active components and their relations through observing these pathways in "Blood Stasis Syndrom" models.The study would reveal scientific connotation of relations between Safflower and Frankincense and provide novel approaches for improving quality of Chinese Potent Medicine and theories of TCMs.

迄今人们无法揭示中医药 "相须""相使"配伍科学内涵。血瘀为"万病"之源，以红花-乳香药对为核心组成的数百方剂在"血瘀证"临床治疗中应用广泛，疗效确切（仅"步长脑心通"年销已达20亿）。但红花-乳香活血抗炎抗氧化"相须""相使"药效物质及相互作用机制不明。前期，我们在国科金（No.81173514）中，用首创的"药效差示血清色谱法"研究发现，红花和乳香的主要组份之中羟基红花黄色素A和乳香酸，均是通过调控NO-cGMP、 NF-κB与Nrf2实现活血抗炎抗氧化效应, 故推测，红花-乳香药对活血抗炎抗氧化"相须""相使"的机制可能是通过协同增效调控NO-cGMP、NF-κB与Nrf2而产生。本课题拟在血瘀证机体，以上述信号通路调控为药效导示，揭示红花-乳香药对配伍的药效物质及其"相须""相使"的内在相互作用机制，为中药大品种质量提升和二次开发、为传承和弘扬中医药理论开创新思路和方法。

迄今人们无法揭示中医药 “相须”“相使”配伍科学内涵。血瘀为“万病”之源，以红花-乳香药对为核心组成的数百方剂在“血瘀证”临床治疗中应用广泛，疗效确切。但红花-乳香活血抗炎抗氧化“相须”“相使”药效物质及相互作用机制不明。.本课题拟在血瘀证机体中揭示红花-乳香药对配伍的药效物质及其“相须”“相使”的内在相互作用机制。1. 确定了红花乳香抗脑缺血再灌注的最大和最小药效时间点，以药效差示血清色谱法筛选出了红花乳香中的五种药效物质，分别是HSYA，α-BA，β-BA，KBA，AKBA。2. 分别对这些活性物质进行体内外作用机制研究，结果表明，在大鼠脑缺血再灌注损伤模型中，KBA和AKBA均可显著减少脑梗死体积，改善神经功能评分，增加SOD等抗氧化酶的水平，激活脑组织中 Nrf2/HO-1通路。α-BA 可抑制寒凝血瘀大鼠血清中多种炎性因子表达，激活NF-kB。β-BA可保护寒凝血瘀证大鼠血管内皮完整性，增加颈动脉血管内皮细胞中的NO / cGMP水平。为进一步增强AKBA 的脑保护靶向作用，我们构建并研究了AKBA羟甲基壳聚糖纳米剂型（AKBA-NPs）。结果显示，AKBA-NPs显著增加了药物的生物利用度及脑靶向性。AKBA-NPs细胞保护作用显著提高，同时可显著减少脑梗死面积，改善神经功能评分，降低细胞凋亡百分比。3. 相关拓展研究发现，AKBA通过TGF-Smad3通路调控高血压重构，α-BA可通过激活MMP-VEGF通路发挥胃保护作用。红花乳香协同作用研究发现，红花中HSYA和AKBA可“相须、相使”发挥抗氧化损伤和血管内皮保护作用。4. 利用基因组，对血瘀证的病理生理机制和红花-乳香药对抗血瘀证的药理作用网络进行了研究。.本项目利用药效差示血清色谱法阐明红花-乳香药效物质，以NO-cGMP， NF-κB 和 Nrf2 所调控的改善微循环障碍，抗炎，抗氧化损伤的信号分子和相关指标为药效导示，深入探讨其“相须”、“相使” 配伍的科学内涵， 为中药方剂、 药对的药效物质基础研究提供了行之有效的研究策略，为传承和弘扬中医药理论开创新思路和方法。