基于胃肠吸收的多维信息融合研究良附丸中药物炮制机理

Compound formulae and processing are the biggest two characteristics of Traditional Chinese Medicine, which can be symbols differing from other medical systems. In our earlier study supported by National Natural Science Foundation (81303227), we found that the absorption difference of Cyperi rhizoma’s characteristic components before and after processed by vinegar in gastrointestinal phase was the significant cause of its efficacy, which provide a scientific explanation for its vinegar processing leading efficacy to the liver from the level of a single medicine. Thus, the hypothesis of the synergistic effect of drug effect components in the absorption of gastrointestinal tract after processing is the key to increase the effect of Liangfu pills is put forward. Liangfu pill, which is a basic compound used by Chinese medicine to treat clinical epigastric pain caused by of cold and qi stagnation, was chosen as the study object. Firstly, basing on specialties of the clinic syndrome differentiation, an animal model with epigastric pain caused by cold and qi stagnation should be established. Secondly, taking the difference of gastrointestinal absorption as the core, techniques and methods like UPLC-DAD-Q-TOF-MS/MS and Gastrointestinal absorption kinetics would be used to obtain the difference among efficacy expression, components in vitro and absorption in vivo. At last, multidimensional spatial triangular area method, which is consistent with the Chinese Medicine’s characteristics of multi-components and multi-targets, would be used to analyze the data, so the processing mechanism of enhancing the efficacy of medicines in Liangfu pill could be explained. By doing these, not only the processing mechanism of vinegar Cyperi rhizoma could be further enriched and perfected from the level of Chinese Medicine compound formulae, but also the foundation for reasonable clinical application and second modernization developing of traditional Chinese compound formulae could be established.

复方入药和中药炮制是中医临床用药两大特色，是区别其他医疗体系的重要标志。课题组前期在国家自然科学基金（81303227）研究中发现香附醋制前后特征性成分在胃肠吸收环节的差异性是炮制增效重要原因，从单味药层面初步阐明其“醋制入肝”科学内涵。现提出“炮制后药效成分在胃肠吸收环节的协同作用是良附丸中药物炮制增效的关键”的假说，选择中医临床治疗寒凝气滞胃脘疼痛的基础方“良附丸”为研究对象，通过构建符合中医证候寒凝气滞胃脘痛大鼠模型，以“胃肠吸收差异”为核心，采用UPLC-DAD-Q-TOF-MS/MS、胃肠吸收动力学等技术研究比较药物炮制前后组方的“体外成分-体内吸收-药效表征”差异，运用符合中药复方多成分、多靶点特点的多维空间信息融合三角形面积法进行数据拟合，诠释复方“良附丸”中药物炮制增效机理，进一步从中药复方层面丰富和完善香附“醋制入肝”机理，为中药传统制剂的临床合理应用与二次开发奠定基础。

良附丸出自谢元庆《良方集腋》，由高良姜、香附二味中药等分组成，具有温胃理气之功效，是临床治疗寒凝气滞胃脘疼痛的基础方。《良方集腋》记载良附丸：“高良姜酒洗七次，香附子醋洗七次……”。可见原方中高良姜需酒制，香附需醋制。但历版《中国药典》收载良附丸中高良姜均为生品，仅香附需醋制，且其在复方良附丸中炮制机理尚不清楚。.本项目以良附丸为研究对象，通过构建符合中医证候寒凝气滞胃脘痛大鼠模型，比较高良姜和香附炮制前后分别组方后“温胃理气”作用，结果以酒高良姜和醋香附配伍后效果最佳，与《良方集腋》记载一致，印证了古人遣方用药的正确性；以药效为导向，采用系统溶剂法筛选良附丸发挥药效物质基础，结果表明良附丸石油醚部位对寒凝气滞胃脘痛大鼠模型“温胃理气”药效作用最佳，其中以高良姜素、香附稀酮等为其发挥药效主要成分；最后从胃肠吸收和血清代谢组学层面探讨了良附丸“温胃理气”作用机理。大鼠在体肠吸收结果表明，良附丸药效指标成分在十二指肠、空肠和回肠吸收无显著差异，说明胃肠吸收差异可能不是良附丸方中药物炮制前后发挥药效差异机理；进一步比较炮制前后分别组方对模型大鼠血清代谢组学差异，结果表明与空白组相比，寒凝气滞胃脘痛大鼠103个标志代谢物发生显著变化，影响氨基酸代谢、碳水化合物代谢、脂质代谢、核苷酸代谢、辅助因子和维生素的代谢等32条具体通路。良附丸各给药组对差异代谢物数目、代谢物通路均有一定调节作用，其中以D组（酒高良姜+醋香附）>C组（生高良姜+醋香附）>B组（酒高良姜+生香附）>A组（生高良姜+生香附），说明高良姜酒炙和香附醋炙品配伍组方更加有利于恢复因造模对机体的影响，高良姜、香附均炮制后入复方将是良附丸的最佳配伍方式，也从代谢组学层面进一步印证了古人描述良附丸之高良姜应当“酒洗”后入药的科学性。此外，本项目还在药典基础上提升了香附和高良姜药材标准，其中香附质量标准正积极和澳门大学申报欧洲标准。