【作者】 尉小慧；

【导师】 孙秀梅；

【作者基本信息】 山东中医药大学 ， 中药学， 2003， 博士

【摘要】 目的：以麻杏石甘汤为例，进一步探讨中药复方“半仿生提取法（简称SBE法）”研究的模式和意义。方法：以盐酸麻黄碱、氢氰酸、钙离子、甘草次酸、干浸膏为指标，用均匀设计法优选SBE法的工艺条件，并对方中4味药材的15种组合方式、澄清工艺进行筛选；依此条件对SBE法、SBAE法（半仿生提取醇沉法）、WE法（水提取法）、WAE法（水提醇沉法）4种方法提取液的成分、药效、兔血清指纹图谱进行比较。在此基础上，制备麻杏石甘喷雾剂，并对其质量标准、HPLC指纹图谱、稳定性进行考察。结果：麻杏石甘方药SBE法的工艺条件为：3煎用水pH依次为2.0、6.5、9.0，煎煮时间依次为2h、1.5h、0.5h；方药较佳组合方式为麻黄与石膏、甘草与苦杏仁分别煎提再合并；60％醇沉为制剂的较佳澄清工艺；4种方法提取液中指标成分综合评价值Y值顺序为：Y_SBE液＞Y_WE液＞Y_SBAE液＞Y_WAE液；4种方法提取液均能明显延长氨水刺激引起的小鼠咳嗽潜伏期，减少小鼠的咳嗽次数，提高小鼠单核巨噬细胞对碳粒的吞噬功能，对哮喘模型大鼠有增加体重、延长咳喘潜伏期、恢复机体的白细胞数目及正常分布、降低血中及肺组织中内皮素含量、改善支气管肺部病理形态学结构等作用，但对小鼠免疫器官的重量无明显影响，4种方法提取液药效强度无明显差异；兔血清HPLC、HPCE指纹图谱均显示，SBE液在体内的吸收代谢速度整体快于WE液。制备麻杏石甘喷雾剂，初步规定该制剂的质控方法和标准，其指标成分的含量范围分别为：盐酸麻黄碱不得少于1.2mg·mL^-1，氢氰酸含量范围为0.7mg·mL^-1～1.1mg·mL^-1，钙离子不得少于34mg·mL^-1，甘草次酸不得少于0.66mg·mL^-1，干浸膏得率不得少于0.24％。初步确定13个峰为制剂HPLC指纹图谱的特征峰，选择其中1峰为参比峰，各特征峰相对保留时间（相对峰面积）依次为：0.31（0.50），0.35（0.21），0.44（0.42），0.53（0.91），0.58（0.95），0.86（0.21），0.90（0.42），0.96（0.36），1.07（0.21），1.22（0.56），1.50（0.38），1.58（0.10）。对其稳定性考察结果显示，该制剂有效期为1年。结论：SBE法能够遵循中医药整体理论指导，体现中药复方多靶点、多环节、多途径的作用特点，是较常规WE法更优的提取新方法，可为中药新药研究中的药效物质提取提供新的研究思路和方法，具有较高的科学性和实用性。更多还原

【Abstract】 Objective: To approach the basic mode and the meaning of the compound prescriptions of the traditional Chinese medicine (TCM) extracted by "the semi-bionic extraction method (SBE method)" with Maxing Shigan Tang as the illustration.Method: With hydrochloric ephedrine> glycyrrhetinic acid^ hydrocyanic acid> calcium-ion and dried extract as the indexes, the uniform-design was used to optimize the prescription’s extract conditions for the SBE method. The optimal combination for the four medical herbs and the optimal clarify conditions for the prescription were selected. According to the former conditions, the 4 kinds of extract methods, including SBE (semi-bionic extraction) method, SBAE (semi-bionic extraction and alcohol precipitate) method, WE (water extraction) method and WAE (water extraction and alcohol precipitate) method, were compared in the contents of the index ingredients, in the pharmacologic activity and in the fingerprinting of the rabbit’s blood serum. Based on the former study, the Maxing Shigan aerosol was prepared and the quality control standards, the HPLC fingerprinting, and the stability of the preparation were studied.Results: The conditions of the SBE method are that the pH values of the water for the 1st, 2nd and 3rd decoctions are 2.00, 6.50 and 9.00; and the times of the 3 decoctions are 2.0h, l.Oh and 0.5h. The optimal combination of the prescription is Ephedra and Gypsum, Liquorice and Bitter Apricot Seed decocted respectively and the decoctions combined finally. The optimal clarify condition is 60% alcohol sediment. The order of the total index Y value of the five ingredients is YSBE>YWE >YSBAE>YWAE.The 4 kinds extractions can prolong the cough incubation period of the mousestimulated by aqua ammonia, decrease the cough times and improve the mononuclear-macrophage’s phagocytic function of the mouse obviously. The 4 kinds of extractions can also enhance the weight, prolong the cough incubation period, restore the leukocytes’ count and classification, reduce the ET-1 content in the blood and the lung tissue, and improve the bronchi’s and the lung’s pathological structure of the asthma cat, but have no effect on the weight of the mouse’s immune organs. With statistics procession there have no significant difference among the 4 kinds of extractions. The HPLC fingerprinting and the HPCE fingerprinting of the rabbit blood serum show that the SBE decoction’s metabolic rate is more rapid than the WE decoction in vivo.Maxing Shigan aerosol was prepared. The quality control standards were formulated and the content range of the index ingredients are: hydrochloric ephedrine above 1.2 mg ?ml/1, hydrocyanic acid form 0.7mg ?ml/1 to l.lmg ?ml/1, calcium-ion above 34 mg ?ml/1, Glycyrrhetinic acid above 0.66mg ?ml/1 and dried extract above 0.24%. 13 peaks are established as the characteristic absorption peaks of the preparation’s HPLC fingerprinting, the relative retention time (the relative peak area) of the characteristic peaks in order are 0.31(0.50), 0.35(0.21), 0.44(0.42), 0.53(0.91), 0.58(0.95), 0.86(0.21), 0.90(0.42), 0.96(0.36), 1.07(0.21), 1.22(0.56), 1.50(0.38), 1.58(0.10). The study of the preparation’s stability shows that the expiration date of the aerosol is 1 year.Conclusion: SBE method can persist the holism theory of TCM and embody the multi-target, multi-link and multi-channel features of the traditional Chinese medicine’s action. It is a new extract method superior to the conventional WE method. SBE method provides new thinking and new method for the study of the prescriptions of the TCM and has high scientific meaning and high practicability.更多还原