【作者】 魏英勤；

【导师】 袁久荣；

【作者基本信息】 山东中医药大学 ， 中药学， 2003， 博士

【摘要】 中药复方（又称方剂）是中医遣方用药的主要形式，中药复方化学的研究又是制约中药复方研究的“瓶颈”所在，弄清中药复方产生疗效的物质基础，对于新药的研制开发，中医理论方剂配伍内涵的科学阐明，对于生命科学的深层次认识，以及中医药现代化都具有重要意义。本文在中药复方研究的基础上，结合实践体会，认为有效部位是中药复方研究的核心问题，提出了“有效成分组是采用现代各种分离手段获得的能够保持原中药复方临床疗效的不可继续分割的分子集合体”的观点，并且认为有效部位的深入细致的研究，将为最终实现中医药理论从药味配伍向化学成分配伍的质的飞跃奠定基石。本课题选用主要由黄连、黄柏等药材组成的经临床验证疗效确切的中药复方血糖宁为研究对象，以降血糖效果为评价指标分离分析了血糖宁的有效部位。主要工作及成果如下： 建立了同时测定血糖宁复方原料药材、原方提取物中四种生物碱含量的RP-HPLC方法，并以四种生物碱、总碱和浸膏得率为指标采用正交设计优化了血糖宁的提取工艺。采用HPCE法比较了喷雾干燥法和烘干法对血糖宁原方提取物有效成分和性质的影响。 以大孔树脂洗脱液中生物碱的含量作指标，选择洗脱液乙醇浓度、洗脱液用量、洗脱液盐酸浓度三个因素，用L₉（3⁴）正交表优化了黄连提取液的大孔树脂洗脱条件。并以洗脱液中生物碱的含量作指标，考察了上样体积、上样量、上样流速、清洗溶剂对于黄连提取液吸附效率的影响。分离了血糖宁的大孔树脂洗脱部位并进一步进行了分离纯化与含量测定。 通过控制剂量、禁食时间等因素，使制作的四氧嘧啶糖尿病小鼠模型造模成功率可达100％，死亡率为0，糖尿病小鼠30日之内稳定。通过药理实验确认了中药复方血糖宁的有效部位，进行了血糖宁部位Ⅲ₂的主要药效学实验的研究，考察了影响血糖测定的因素，并测定了血糖宁部位Ⅲ₂的LD₅₀。 进行了血糖宁的质量标准研究，采用TLC对方中主要组成药味及制剂进行了定性鉴别，利用TLCS和HPLC对方中主要组成药味及制剂中的生物碱进行了含量测定，并并考察了其初步稳定性。 建立了血糖宁原料药材黄连、部位Ⅲ和部位Ⅲ₂的HPCE指纹图谱，并将其用于质量控制和评价。建立了血糖宁及其缺味药部位＊的Bayes法识别模型，该模型对血糖宁及其缺味药部位＊正确识别率为100％。进行了药动学方面的初步研究，建立了灌胃黄连煎剂的大鼠尿样中成分的eCE指纹图谱，发现黄连生物碱主要以原型药物排出，与文献结论一致。 本文分离并明确了中药复方血糖宁的有效部位，并对其进行了含量测定、药效、质量控制、指纹图谱等方面的研究，研究结果表明生物碱是中药复方血糖宁降血糖作用的主要物质基础。实验结果将形成中药新药的有关资料，从而产生较大的社会效益和可观的经济效益，同时也将为血糖宁有效成分组的全面揭示、为中医用药组方机理和规律的深层次认识提供素材和实验依据。更多还原

【Abstract】 Chinese herbal formulae also named as recipe are the main means to treat disease by doctor of traditional Chinese medicine and the study on chemical component of Chinese herbal formulae are the key problem to research Chinese herbal formulae. To understand the substance producing theroputic effect fully is of significance to explore new drugs, to explain the mechanism of medicines compatility and to deepen the cognition of biological science. In this paper, the effective fraction was thought as the core of study on Chinese herbal formulae, and a new viewpoint was proposed, i. e, the group of effective components is the smallest molecule group obtained by modern separation means and still maitaining the theroputic effect of original Chinese herbal formulae. In addition, it was also thought that penetrating into the effective fraction would lay a foundation for realizing the deepening of TCM theory from medicines compatility to chemical components compatility. On the bases of these viewpoints, a Chinese herbal formula---Xuetangning was sysmetically studied to separate the effective fractionguided by the hypoglycemic effect. Main works are as follows:Firstly, the RP-HPLC determination method of alkaloids in Xuetangningwas established, and the extraction process of Xuetangning was optimized?using orthogonal experimental design. In addition, HPCE was used to compare the influence of different drying method on the components and characterinof Xuetangning. In the second chapter, the elution conditions of Rhizoma Coptis extraction from macroporous resin were optimized using L9(3)4 orthogonal experimental design and the influencing factors on the absorption efficiency was explored. In addition, Xuetangning fractions desorpted from macroporous resin were prepared and analysed. In the third chapter, alloxan diabetic mice model was made and the influencing factors on the succeeding rate such as dosage of alloxan and fasting time were explored and the effective fraction of Xuetangning. was verified. In the forth chapter, the quality standard of Xuetangning capsule was drawn up, and the stability of drug material and Xuetangning was studied. In the last chapter, the finger-printings of Coptis chiensis Franch, fraction III and fraction III2 of Xuetangning preparation were made by HPCE and the Bayes identification model which could identify fraction III of Xuetangning and its modified formulae accurately was established. The finger-printing of components in urine samples of rat after administration with Coptis chiensis Franch decocta was also studied.In a word, the effective fraction of Xuetangning was verified, and its determination, finger-printing, theroputic effect and quality control were studied thoroughly in this paper. Results showed that alkaloids are the main material base for descending serum glucose in alloxan diabetic mice. The data of experiment would evolve the relevant materials for new drugs, so considerable social and economic benefit would be created. At the same time these data would provide raw materil and fresh evidence to reveal the group of effective components and to deepen the cognition of TCM compatil i ty mechenism.更多还原