神经营养因子基因在神经干细胞中的翻译与表达  

A primary study on the transcription and expression of neurotrophin genes in neural stem cells

作　　者：孙崇然[1] 王忠诚[1] 李晋[1] 张建民[2] 王雅杰[3] 张涵[1] 安沂华[1] 

机构地区：[1]首都医科大学北京市神经外科研究所,北京100050 [2]浙江大学医学院附属第二医院神经外科 [3]首都医科大学附属北京天坛医院检验中心

出　　处：《中华实验外科杂志》2008年第8期962-963,共2页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30371452）;北京市自然科学基金资助项目（7042014）;北京市科技新星计划项目资助项目（H020821360130）

摘　　要：目的检测体外培养的神经干细胞三种神经营养因子基因的翻译水平和神经生长因子的分泌。方法取孕16d胚胎SD大鼠皮层细胞，无血清选择性培养液培养神经干细胞，酶联免疫吸附试验（ELISA）法检测第1-7代细胞体外分泌神经生长因子的速度，相对定量实时定量聚合酶链反应（real time-PCR）检测第5-7代细胞脑源性神经营养因子、神经生长因子、神经营养因子-3 mRNA的相对表达量。结果体外培养的神经干细胞在第6代分泌NGF速度最大，（17．80pg／10^6细胞·d^-1）。在第6代BDNF和NT-3基因的相对翻译量最大，在第7代NGF的相对翻译量最大（与对照比值分别为29．446、35．017和16．220）。结论神经干细胞在体外翻译、表达并分泌神经营养因子。Objective To investigate the relative transcription rate of neurotrophins genes and the expression rate of nerve growth factor （NGF） secreted by neural stem cells. Methods The neural stem cells were harvested from the cortex of E-16 Sprague-Dawley rats, cultured in the neural stem cell-conditioned Dulbecco＇ s Modified Eagle＇ s Medium （DMEM）. The secretion rate of NGF in the cells of 1st -7th passages was determined by ELISA assay, and the relative translation rate of brain-derived neurotrophic factor （BDNF）,NGF and neurotrophin-3 （NT-3） in the cells of 5th -7th passages were determined by partial quantitive real-time PCR. Results The peak of NGF secretion rate was at the 6th passage （ 17.80 pg/10^6 cells ·day） in neural stem cells. The peaks of translation rate of BDNF and NT-3 were at the 6th passage （29. 446 and 35. 107 ）, and of NGF, at the 7th passage （ 16. 220）. Conclusion Neural stem cells translate, express and secrete neurotrophins in vitro.

关 键 词：神经干细胞 神经生长因子 脑源性神经营养因子 基因表达 

分 类 号：R741[医药卫生—神经病学与精神病学]