续断药材发汗加工过程的酶促作用和化学反应机制研究

Sweating is a unique process for the initial processing of Chinese herbal medicines and an important link to the quality of medicinal herbs. During the sweating process, the secondary metabolites in the tissues may undergo a series of reactions such as enzymatic and chemical transformations, resulting ultimately in the unique quality characteristics of medicinal materials. therefore, we can reveal the quality changes essence after sweating of medicinal herbs by elucidating the mechanisms of enzymatic and chemical transformation. Dipsaci Radix is a Chinese herbal medicine that is definitely stipulate to sweat processing after harvesting in the Chinese Pharmacopoeia. Studies have shown that the chemical composition and species changes significantly after sweating of Dipsaci Radix, however, the connotation of related quality changes has not yet been scientifically explained. This project reveal the reaction mechanism that the chemical conversion in the enzymatic action and triterpene saponins and cyclic glycosides in the sweating process, using synthetically these technologies such as proteomics and chemical composition analysis, and clarify the scientific connotation that the quality and color changes after sweating of Dipsaci Radix. And based on the above basics, its further summarizing the qualitative change common regularity and common research techniques in the sweating process of herbal medicine. It is of great significance to establish standards for the initial processing of standardized herbal sweats and guarantee the quality of medicinal materials.

发汗是中药材初加工的独特工艺，也是影响药材品质的重要环节。中药材在发汗处理中，组织内的次生代谢物质可能会发生一系列酶促和化学转化等反应，最终形成药材独特的品质特征，通过阐释其反应机制可以揭示导致药材发汗后质量变化的本质。.续断是中国药典明确规定采收后需发汗加工处理的中药材。已有研究表明续断发汗加工后次生代谢产物的含量及种类变化明显，但相关的质量变化内涵尚未得到科学阐释。本项目综合运用蛋白组学、化学成分分析等技术，揭示续断发汗过程中的酶促作用和三萜皂苷、环烯醚苷类化学转化的反应机制，阐明续断发汗后质量和颜色变化的科学内涵，并在此基础上进一步总结中药材发汗过程中质量变化的共性规律，形成共性研究技术，为建立规范的药材发汗初加工标准，保障药材品质具有重要意义。

中国药典规定中药材续断作为临床饮片需要经过“发汗”的炮制加工。本项目运用蛋白组学和化学成分分析等技术，揭示了续断药材传统发汗加工的科学内涵。通过四年的研究工作，取得的重要结论是：续断发汗加工后，药材根断面的颜色较发汗前加深是根中环烯醚萜类成分发生转化所致，而环烯醚萜苷类成分之所以发生转化是植物中的过氧化酶POD和SOD活性作用。在发汗过程中，以川续断皂苷Ⅵ为代表的成分含量升高主要由CYP72A和UGT酶主导，其中DaCYP72A61家族蛋白酶对其化合物变化发挥显著作用，DaCYP72A61v1蛋白能够将齐墩果酸转化为其它三萜类物质，上述反应是酶促作用下的氧化还原反应。此外，续断发汗过程还存在非酶促反应，植物中的共生真菌中，镰刀菌属真菌具有促进川续断皂苷生成的作用。本项目以续断药材为研究对象，揭示了皂苷和环烯醚萜苷类为主要成分的中药材发汗的共有机制，这为后续形成“发汗加工”共性关键控制技术，建立规范、统一、标准的药材发汗初加工方法，保障药材品质具有重要意义。