基于体内外成分转化的丹参酒炙增效炮制原理研究

Radix Salviae Mil Tiorrhizae has become one of the largest amounts among Traditional Chinese Medicines in China. The effects of its processed product in activating blood and eleminating stasiswere were significantly enhanced, but the synergistic mechanism has not been clear. Previous study discovered that the components of this medicinal material changed significantly after wine processed. It is postulated that the synergistic mechanism comes from the transformations of original components, which become new components with more activities easier for the body to absorb. This project focuses on the changed components during the wine processing and plans to discover the target components from the raw and prepared medicinal materials in the plasma of rats by UPLC-Q-TOF-MS detecting, principal components comparison and partial least squares-discrimi-nant analysis. These components will be enriched and isolated by various chromatographic methods, while their structures will be elucidated by modern spectroscopic ways. After that, transformation regulations of the obtained compounds will be studied by modem analysis technologies and analog processing technology. At last, the internal transformation rule and the intestinal absorption differences of these components will be investigated by using in situ perfusion method in rats. This project may find out the bioactive components of wine prepared radix salviae miltiorrhizae, clarify the synergistic mechanism and provide scientific basis for establishing the processing technics regulation and quality control. It is also helpful for the researches on the wine prepared herbal materials and the development of Traditional Chinese Medicine.

丹参是我国年用量最大的中药品种之一，酒炙后活血祛瘀的功效显著增强，但酒炙增效的机理尚未得到科学解释。课题组前期研究发现丹参酒炙前后成分的结构和含量变化显著，据此推测丹参酒炙增效机理是原有成分转化成更利于机体吸收、活性更强的成分。本项目拟以丹参酒炙前后质变、量变成分为目标化合物，运用UPLC-Q-TOF-MS联用等技术研究其生、炙品化学成分谱和大鼠血浆移行成分谱，以主成分分析和偏最小二乘-判别分析建立谱-谱相关性数学模型，发现目标化合物。再综合运用各种分离手段对目标化合物进行富集、分离，应用化学方法和波谱方法鉴定其结构。然后通过现代分析技术和单体成分模拟炮制技术，研究单体成分的转化规律。最后通过在体肠灌流模型探讨成分的体内转化规律和肠道吸收差异。本项目有助于揭示酒炙丹参的药效物质基础，阐明酒炙增效机理，为规范其炮制工艺、制定质量标准提供依据，为研究酒炙在中药炮制和中医药发展中的作用提供帮助。

本课题以丹参酒炙前后质变、量变成分为目标化合物，运用HPLC 技术比较分析丹参不同炮制品指纹图谱差异，将丹参素、原儿茶醛、咖啡酸、迷迭香酸、丹酚酸B、紫草酸、二氢丹参酮I、丹参酮IIA、丹参酮I、隐丹参酮等成分作为目标化合物进行研究。对目标化合物进行富集、分离、鉴定，共分离、鉴定出12个化合物，其中水溶性成分7个，脂溶性成分5个，水溶性成分分别是原儿茶醛、咖啡酸、阿魏酸、迷迭香酸、紫草酸、丹酚酸B、丹酚酸C，脂溶性成分分别是、二氢丹参酮I、隐丹参酮、丹参酮I 、丹参新酮、丹参酮IIA。.通过现代分析技术和单体成分模拟炮制技术，研究单体成分体外的含量变化和转化规律，发现丹参素、原儿茶醛、咖啡酸、迷迭香酸等水溶性成分和二氢丹参酮I、丹参酮IIA、丹参酮I等脂溶性成分含量升高，丹酚酸B、紫草酸、隐丹参酮的含量降低。对迷迭香酸、丹酚酸B、隐丹参酮、二氢丹参酮I、丹参酮IIA进行模拟炮制，采用UPLC-EQ/MS分析鉴定，结果表明，上述5种主要成分在加水（黄酒）、加热的条件下发生分解，生成了丹参素、阿魏酸、咖啡酸、紫草酸、二氢丹参酮I等成分。利用大鼠在体胃肠灌流模型，采用HPLC法比较丹参酒炙前后5 种质变与量变特征化合物（迷迭香酸、丹酚酸B、隐丹参酮、二氢丹参酮I、丹参酮IIA）大鼠小肠的吸收差异及在大鼠体内的代谢产物。结果发现，在小肠内各药物吸收百分率为：原儿茶醛＞丹参素＞咖啡酸＞阿魏酸＞迷迭香酸＞丹酚酸B＞紫草酸，吸收速率常数：原儿茶醛＞阿魏酸>咖啡酸＞丹参素>迷迭香酸>丹酚酸B>紫草酸。大分子化合物丹酚酸B、紫草酸、迷迭香酸在大鼠小肠内可转化为原儿茶醛、丹参素、咖啡酸、阿魏酸。.结果表明，丹参中迷迭香酸、丹酚酸B、隐丹参酮、二氢丹参酮I、丹参酮IIA 5种主要成分的体内外转化规律一致，均生成了丹参素、阿魏酸、咖啡酸、紫草酸、二氢丹参酮I等分子量小、较其原型更利于被机体吸收的活性成分，从而达到酒炙增强活血化瘀药效的目的。