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作 者:朱俊平[1,2] 李龙 曾燕[1,3] 范诗琪 梁慧慧[1,2] 何红 张喜利 贺福元 刘文龙[1,2] ZHU Jun-ping;LI Long;ZENG Yan;FAN Shi-qi;LIANG Hui-hui;HE Hong;ZHANG Xi-li;HE Fu-yuan;LIU Wen-long(Pharmacy College,Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Key Laboratory of Druggability and Preparation Modification for Traditional Chinese Medicine,Changsha 410208,China;Second Affiliated Hospital of Shaoyang Academy,Shaoyang 422000,China;Hunan Children's Hospital,Changsha 410007,China)
机构地区:[1]湖南中医药大学药学院,湖南长沙410208 [2]中药成药性与制剂制备湖南省重点实验室,湖南长沙410208 [3]邵阳学院附属第二医院,湖南邵阳422000 [4]湖南省儿童医院,湖南长沙410007
出 处:《中国中医药信息杂志》2018年第9期61-65,共5页Chinese Journal of Information on Traditional Chinese Medicine
基 金:国家自然科学基金(81270055);湖南省教育厅科研优秀青年项目(15B174)
摘 要:目的优选黄连蛋白质最佳提取方法,比较分析野生与种植黄连不同部位的蛋白质差异。方法采用聚丙烯酰胺凝胶电泳和考马斯亮蓝法,以蛋白质条带丰度、蛋白质相对分子量、每克样品中蛋白质含量为指标,比较3种提取方法(水提法、Tris-HCl法、硫酸铵沉降法)所得黄连蛋白质的差异,并对野生与种植黄连不同部位蛋白质的差异进行分析与评价。结果优选得出黄连蛋白质最佳提取方法为Tris-HCl法,所得黄连蛋白质丰度最高,且每克药材中蛋白质含量最高。野生与种植黄连不同部位的蛋白质有明显差异,蛋白质丰度及每克药材中蛋白质含量均为:种植黄连根茎>野生黄连根茎>种植黄连茎叶>野生黄连茎叶>种植黄连须根。聚类分析结果显示,野生与种植黄连根茎蛋白质关联关系较为显著。结论本研究优选的Tris-HCl法可充分提取黄连中的蛋白质;黄连根茎蛋白质含量明显高于其他部位。Objective To optimize the extraction method of Coptidis Rhizoma protein; To compare and analyze the differences of protein in different parts of wild and cultivated Coptidis Rhizoma. Methods SDS-PAGE gel electrophoresis and Coomassie brilliant blue method were used to compare the differences of the protein of Coptis chinensis from the 3 extraction methods(water extraction, Tris-HCl and ammonium sulfate precipitation), and the protein in different parts of the wild and cultivated Coptis chinensis, and analyzed and evaluated the differences. Results The optimum extraction method of Coptis chinensis protein is Tris-HCl method, and the protein content of Rhizoma Coptidis is the highest, and the content of protein is the highest in every gram of medicinal material. The differences among different parts of wild and cultivated Coptidis Rhizoma were obvious, and the ranks for protein abundance and protein content per gram were: rhizome of cultivated Coptis chinensis rhizome of wild Coptis chinensis stem and leaf of cultivated Coptis chinensis stem and leaf of wild Rhizoma Coptidis fibrous roots of cultivated Coptidis Rhizoma. Cluster analysis showed that the correlation between protein of wild and cultivated rhizome of Coptidis Rhizoma was obvious. Conclusion The optimum Tris-HCl method can extract the protein from Coptidis Rhizoma, and the protein content of roots of Coptidis Rhizoma is significantly higher than other parts.
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