【作者】 吕鹏月；

【导师】 石俊英；

【作者基本信息】 山东中医药大学 ， 中药学， 2004， 硕士

【摘要】 目的：紫河车是人的胎盘经炮制加工而成的贵重滋补药，具补气养血、益肾补精之功。目前，随着我国计划生育工作的深入开展，紫河车的药材资源越来越少，而紫河车的多剂型的开发又扩大了对市场的需求。因此，为紫河车寻找代用品已成为当务之急。猪、牛、羊胎盘均来源于哺乳动物胎盘，且资源丰富，是紫河车的理想代用品，目前国内市场上有用猪、牛、羊胎盘混淆品代替紫河车。鉴于此，本课题立项进行了“紫河车及其代用品的药理学与质量评价研究”。旨在探讨猪、牛、羊胎盘代替紫河车应用的可行性，探讨紫河车及其代用品的药理学作用机制，为初步建立紫河车与猪、牛、羊胎盘的鉴别方法与质量控制指标提供科学依据。方法：本课题主要进行了紫河车及其代用品猪、牛、羊胎盘的鉴别研究、主成分分析、药理学研究、质量评价方法与标准研究。鉴别研究进行了理化鉴别、显微鉴别、薄层鉴别、聚丙烯酰胺凝胶电泳鉴别和傅立叶变换红外光谱法鉴别；主成分分析进行了蛋白质、氨基酸和微量元素的测定，分别采用凯氏定氮法、氨基酸分析法和电感耦合等离子体发射光谱法；主要药效学研究进行了小鼠的补血、学习记忆、抗应激能力和免疫能力的研究；急性毒性研究进行了小鼠灌胃给药的最大给药量测定法；质量标准研究进行了鉴别、检查和含量测定。检查项包括水分、总灰分、酸不溶性灰分、重金属限度的测定，含量测定项包括蛋白质含量、水溶性浸出物、醇溶性浸出物的测定。结果和结论：鉴别研究的结果表明，常规的理化鉴别、显微鉴别、薄层鉴别等均难以区分紫河车及猪、牛、羊胎盘。聚丙烯酰胺凝胶电泳鉴别具有显著差异，可以作为紫河车及猪、牛、羊胎盘的鉴别依据。本课题首次采用傅立叶变换红外光谱法进行了紫河车及其代用品的鉴别，实验结果表明紫河车及猪、牛、羊胎盘的普通红外光谱图相近，二阶导数光谱图有差异但不显著，二维红外光谱图有显著差异，可有效、无损地鉴别四种胎盘。紫河车及猪、牛、羊胎盘的氯仿和乙醇提取物的薄层层析图相似、普通红外光谱图相似，表明紫河车与猪、牛、羊胎盘可能具有相似的物质基础。主成分分析的研究结果表明，紫河车与猪、牛、羊胎盘所含蛋白质含量相近，均含有钙、锌、铁三种人体必需的微量元素，只是含量略有不同；结合笔者发表论文对紫河车及猪、牛、羊胎盘的氨基酸测定结果，进一步提示：紫河车及猪、牛、羊胎盘均来源于哺乳动物胎盘，可能具有相似的物质基础。 药效学研究的结果表明，小鼠灌胃给予紫河车混悬液，对失血性贫血和化学损伤性贫血均有明显的治疗和预防作用(P<0 .01)，可显著改善腹腔注射东食若碱所致的小鼠记忆获得障碍和皮下注射亚硝酸钠所致的小鼠记忆巩固障碍(P<O乃1)，可显著增加小鼠负荷游泳时间和在缺氧状态下的存活时间(P<0.01)，可显著增加小鼠免疫器官胸腺和脾脏的重量(P<0.01)。猪、牛、羊胎盘在上述的药效学指标中与对照组比较亦有显著差异(P<0 .01)，与紫河车组相比无显著差异(P>，05)。实验结果表明，在补血、改善学习记忆、抗应激能力和提高免疫力方面，猪、牛、羊胎盘与紫河车药理作用近似。 急性毒性试验的研究结果表明，小鼠灌胃给予紫河车及猪、牛、羊胎盘混悬液的最大剂量均为30叭g，相当于临床人用量的300倍，表明4种样品的毒性小，安全性高。 对紫河车及其代用品的10批样品进行了质量评价的初步研究，包括鉴别、检查和含量测定，认为可考虑将该三项内容增订入紫河车的质量标准，并初步拟订了猪、牛、羊胎盘的质量评价指标。更多还原

【Abstract】 OBJECTIVE To investigate the feasibility of Placenta suis,Placenta bovis and Placenta caprae as substitutions of Ziheche(placenta hominis) and establish suitable quality standards for Ziheche and its substitutions.METHODS The identifications of four mammalian placentaes were investigated by the analysis of Polyacrylamide gel electrophoresis,Fourier Transform Infrared (FTIR) spectroscopy, physical and chemical method , microscopy and TCL method. The contents of nitrogen in four placentaes were determined by Kjeldahl’s method, and trace elements determined by inductively coupled plasma-atomic emission spectrometry(ICP-AES). The pharmacodynamic studies involves the nourishing blood effect,learning and memory effect,anti-fatigue,anti-anoxia and immune effect. The nourishing blood effect of Ziheche and its substitutions ,the effects of them on hemorrhagic anemia and on anemia induced by chemical damage of cyclophosphamide in mice had been observed.The step down test was used to investigate the effects of four mammalian placentaes on learning and memory impairment induced by scopolamine and sodium nitrite in mice.Anoxia and fatigue animal models in mice were used to reasearch the anti-fatigue and anti-anoxia abilities of four placentaes.The immune organs weight of mice were determined to evaluate the immunological function of four mammalian placentaes.Reasearches on maximal tolerance limit in mice were carried on. The identification, examination and content determination of ten samples of four placentaes were investigated,including the examinations on water content,total ash,acid-insoluble ash and heavy metals,and the determinations on protein,water extract and 75% ethanol extract.RESULTS AND CONCLUSIONS The technique of PAGE is effective on indentifying four placentaes,but not on processed samples. Ziheche and its substitutions were directly determined with FTIR spectroscopy for the first time,and this method is reliable.The contents of protein and trace elements show that Ziheche had similar components with its substitutions,but different in amount.The pharmacodynamic studies showed that compared with Ziheche ,its substitutions all showed the similar significant effects on antianemic and regaining effect on erythrocyte and hemoglobin, alleviating the learning and memory impairment induced by scopolamine and sodium nitrite,improving the ability of anti-anoxia and anti-fatigue,and increasing the spleen index and chest gland index in mice(P>0.05).This indicates that ziheche could be considered to be substituted by other three placentaes on these aspects. The maximal tolerance dose of four placentaes was 30g/kg.This result suggested that the maximal tolerance dose of mice was 300 times as much as that of human. Reasearches on idenfication,examination and content dermination in ten samples of four placentaes would be helpful for establishment and improvement of quality standards.更多还原