基于药效差示血清色谱法-代谢组学技术研究丹参-降香对药治疗冠心病的药效物质及作用机制

Salvia miltiorrhiza-Dalbergia odorifera Jun-Shi coupled-herbs has been widely used for the treatment of coronary artery disease (CAD) for hundreds of years. However, its effective substance and compatible mechanism remain unknown. Our previous study showed that Salvia miltiorrhiza-Dalbergia odorifera could reduce the level of serum free fatty acid and the ratio of acetyl coenzyme A / coenzyme A in myocardium, indicating that the mechanism of Salvia miltiorrhiza-Dalbergia odorifera in the treatment of CAD might be related to the inhibition of myocardial fatty acid oxidation. The prescription of traditional Chinese medicine is the combination of herbs based on "syndrome" according to the holism of traditional Chinese medicine and the process of syndrome differentiation and treatment. However, most prescription researches presently only focus on one aspect of the therapeutic mechanism, ignoring the overall response of the body. Therefore, based on the previous results, according to the metabolomics differences of serum and myocardial tissue between the maximum and minimum efficacy point of treating CAD by Salvia miltiorrhiza-Dalbergia odorifera through Differential Efficacy of Serum Chromatography Analysis (DESCA) combined metabolomics in mini-swines with blood stasis syndrome due to chronic myocardial ischemia, we aim to trace the exogenous effective substance and screen the endogenous metabolites closely related to pharmacodynamics in order to clarify the mechanism of treating CAD by Salvia miltiorrhiza-Dalbergia odorifera through regulating the glucose and lipid metabolism. Our research will further reveal the scientific connotation of the composition of Salvia miltiorrhiza-Dalbergia odorifera and be expected to make up for the weakness in the study of traditional Chinese medicine. Furthermore, we hope to push Salvia miltiorrhiza-Dalbergia odorifera and its composition of prescription internationally through our work.

丹参-降香君-使对药数百年来广泛用于治疗冠心病，但其药效物质和配伍作用机制一直未被阐明。我们的前期研究结果发现，丹参-降香可降低血清游离脂肪酸水平和心肌乙酰辅酶A/辅酶A比值，提示丹参-降香治疗冠心病的机制可能与抑制心肌脂肪酸氧化有关。传统中药方剂是根据中医整体观和辨证论治过程而得出对"证"的方药，但目前大多数方剂的研究仅着眼于治疗机制的某一方面，忽视了机体的整体反应。因此，我们在上述研究基础上，拟建立小型猪的冠心病慢性心肌缺血模型，采用药效差示血清色谱法（DESCA）与代谢组学技术，根据丹参-降香治疗最大、最小药效点血清及心肌组织代谢组的差异，示踪分离出外源性药效物质，并筛选出与药效最密切相关的内源性代谢物群，阐明丹参-降香调节糖脂代谢，治疗冠心病的具体作用机制，进一步揭示丹参-降香配伍的科学内涵，期望弥补中医药研究中的薄弱环节，促进丹参-降香及其组成方剂的国际推广。

丹参-降香君-使对药数百年来广泛用于治疗冠心病，但其药效物质和配伍作用机制一直未被阐明。我们在心肌缺血/再灌注（MI/R）损伤大鼠和慢性心肌缺血小型猪模型中均发现，丹参-降香发挥心肌保护作用的代谢组学特征及作用机制均与抑制心肌细胞对游离脂肪酸的摄取和氧化，促进心肌对葡萄糖的氧化和利用，进而减少缺血心肌氧耗，改善心肌能量代谢密切相关。单用降香亦可抑制线粒体分裂，改善后负荷增加型心衰小鼠的心脏功能。进一步的代谢组学聚类分析结合药效差示血清色谱法（DESCA）研究结果表明，丹参-降香发挥心肌保护作用的可能药效物质为丹酚酸B（SalB）、丹参素、隐丹参酮、丹参酮IIA及紫铆花素（BUT）。对代表性药效物质延伸的研究结果表明，BUT和SalB对糖尿病合并心肌损伤小鼠的保护作用机制分别为：BUT激活AMPK/Akt/GSK-3β通路，促进Nrf2解离入核与抗氧化反应原件结合，上调抗氧化酶的活性，稳定内源性抗氧化系统，减少活性氧调控的心肌细胞凋亡；SalB上调谷氨酰胺酶的表达，促进谷氨酰胺代谢，保护内皮细胞，促进人脐静脉内皮细胞增殖，增强血管生成，改善心脏功能和心脏重塑。在对DESCA-代谢组学技术研究丹参-降香心肌保护的药效物质及机制形成较标准的整体方法学后，对代表性药效物质丹参素的研究结果表明，丹参素磷酸化Akt 保护大鼠内皮祖细胞（EPCs）免受缺氧损伤，激活基质细胞衍生因子-1α及其受体CXCR4生物轴提高EPCs血管生成能力，促进缺血心肌血管新生，进而发挥对心肌梗死大鼠的心肌保护作用。以上研究结果不仅使我们对丹参-降香君-使对药有了新的全面的认识，还为使用代谢组学技术研究中药复方的药效物质及作用机制提供了理论依据和方法学参考。据此发表SCI论文5篇，中文核心期刊论文7篇；培养硕士研究生3名，获评2018年校级优秀学位论文1名，课题负责人获评空军军医大学（原第四军医大学）优秀硕士研究生导师；课题负责人和课题组成员获得共青团中央中国光华科技基金会医学奖学金。