牛脑海马NGF分离纯化及微量元素含量检测  

Separation and Purification of NGF from Bovine Brain Hippocampus and Detection of Trace Elements

作　　者：顾鹏毅[1] 代萌[1] 杨栋[1] 高娜[2] 李宁禾[1] 孙晋民[1] 

机构地区：[1]中国医科大学实验技术中心一部暨国家中医药管理局中药生物工程科研Ⅲ级实验室,沈阳110001 [2]朝阳市卫生学校,朝阳122000

出　　处：《天然产物研究与开发》2012年第12期1757-1760,1790,共5页Natural Product Research and Development

基　　金：辽宁省科学计划项目(2008225021-1);国家发明专利(476108)

摘　　要：将牛脑海马粗提物经盐析、透析、CM-离子交换层析分离纯化得到2个洗脱峰,其中CM2峰含有NGF。利用Western Blot检测结果表明,NGF的分子量约为55 KD和70 KD;PC12细胞法检测其生物学活性,在CM2分离组分作用下,PC12细胞轴突生长效果优于2 ng/mL标准品NGF;原子吸收分光光度法检测脑海马结构微量元素含量,Zn与Cu无差别,Fe与Zn、Cu有差异(P<0.01);CM2组分中Fe与Cu含量无差异,Zn远高于Fe、Cu含量(P<0.01)。The objective of this study was to purify NGF from the crude extract of bovine brain hippoeampus by salting, dialysis and CM-ion exchange chromatography. Two elution peaks, which contains NGF in the CM2 peak, were isolated and purified. The molecular weights of NGF were determined to be around 55 KD and 70 KD using Western Plot. The bi- ological activity of CM2 fractions was tested using PC12 cells. The results showed that its activity against PCI2 cells＇ ax- onal growth was better than 2 ng/mL NGF standard. In addition,with atomic absorption spectrophotometry, testing trace elements the content of Zn and Cu had no difference in bovine brain hippocampus, but the content of Fe was different from Zn and Cu （P 〈 0.01 ）. In CM2 ,there was no difference in the content of Fe and Cu, but the content of Zn was much higher than that of Fe and Cu （P 〈 0.01 ）.

关 键 词：牛脑NGF 分离 纯化 PC12细胞 微量元素 

分 类 号：Q599[生物学—生物化学]