以组分配伍形式研究伤寒论泻下退黄类方配伍规律及效应机制

Study on compatibility rules and effect mechanisms of purgative and jaundice receding prescriptions selected form Shang Han Lun in the form of multi-component combination.Abstract: The project is guided by the TCM holism concept to study the purgative and jaundice receding prescriptions, Yinchenhao Decoction, Zhizi Dahuang Decoction, Dahuang Xiaoshi Decoction, which are selected from Shang Han Lun (Treatise on Febrile Diseases). Yang and jaundice syndrome animal model is used for it fit the etiology and pathogenesis theory of TCM. The prescriptions are made by multi-component combination, for they have relatively clear material constituents. The optimal component combination of Dahuang and Zhizi pair medicines will be screened firstly by complete random design according to effect indicators. Then, the pharmacodynamics, metabolism, and metabonomics will be sequentially studied on the three prescriptions of multi herbs based on the optimal component combination. The experiments will be conducted sequentially from single drugs to pair medicines and then prescription, on whole animals, organ, cell and molecular levels, to elucidate the compatibility rules and effect mechanisms of the purgative and jaundice receding prescriptions by analyzing the large amount of information obtained from the experiments, and the scientific connotation of the purgative and jaundice receding treatment on jaundice diseases could be revealed. The project is the development of a finished National Natural Science Fund project study, with a novel research direction, advanced methods, and is well designed. The expected research results would enlighten the system research of TCM prescriptions, and provide a substantial basis for the new drug development for Yang and jaundice syndrome.

本项目以中医整体观为指导，对《伤寒论》泻下退黄类方茵陈蒿汤、栀子大黄汤及大黄硝石汤进行研究。应用符合中医病因、病机理论的阳黄证动物模型，采取物质组成相对清晰的组分配伍形式，首先根据体内效应指标以完全随机设计筛选出大黄-栀子核心药对最优组分配比，以求得类方研究的共同本底。在此最优组分配比基础上，对加味组成的三首泻下退黄类方进行系统的药效学、体内代谢和代谢组学的序贯研究。试验从单味药-对药-类方三个层面上递进，在整体动物、器官、细胞、分子四个水平上展开，经过分析处理试验给出的大量信息，阐明泻下退黄类方的配伍规律和体内效应机制，从而揭示泻下退黄法治疗黄疸类疾病的科学内涵。本项目是在已结题的国家自然基金项目研究基础上的进一步拓展，研究思路新颖，方法科学先进，方案设计巧妙，预期研究成果将对中医方剂学的系统研究起到启示作用，也为开发治疗阳黄证黄疸的创新药物奠定扎实基础。

本课题围绕课题任务书的要求圆满完成了如下研究内容。本课题考察了3首泻下退黄伤寒类方各组成药味有效组分提取的最佳工艺，进而提取有效组分，建立各组分的多指标控制质量标准；采用α-萘异硫氰酸酯（ANIT）结合湿热环境的方法，制作阳黄证（湿热并重）的大鼠动物模型；以谷草转氨酶（ALT）、谷草转氨酶（AST）、血清总胆红素（TBIL）为指标，采用正交设计法结合多目标优化统计方法筛选确定了大黄栀子药对、茵陈蒿汤、栀子大黄汤、大黄硝石汤的最优组分配比。经药效学验证，大黄硝石汤及茵陈蒿汤组分配伍组优于汤剂；栀子大黄药对与栀子大黄汤与原方汤剂疗效相当；完成了组分配伍大黄栀子药对、茵陈蒿汤、栀子大黄汤、大黄硝石汤在对空白及阳黄模型大鼠的药动学及体内代谢研究，未检测到组分配伍泻下退黄类方中已知原型入血成分，在部分时间点找到了可能的代谢产物；以保肝降酶、利胆退黄、炎性细胞因子等作为体内外效应检测指标，对比泻下退黄伤寒类方的配伍规律及泻下退黄作用机制，研究表明大黄硝石汤及茵陈蒿汤对阳黄证（湿热并重）的模型大鼠疗效显著并疗效相当，栀子大黄药对单用即有效，是发挥泻下退黄作用的核心药对，栀子大黄汤作用最弱。泻下退黄类方的作用机制可能与下列因素有关：①降低的黄疸动物体内ALT、AST、碱性磷酸酶（ALP）、TBIL、直接胆红素（DBIL）、间接胆红素（IDBIL）②改善黄疸动物体内的免疫亢进状态，降低补体C3水平。③减少炎性细胞因子（白介素-6）IL-6、（白介素-2）IL-2释放。本课题是对泻下退黄类方的系统研究，为临床使用泻下退黄类方治疗黄疸（阳黄证）提供科学依据，也为开发治疗阳黄证黄疸的创新药物奠定了基础。