脑皮质星形细胞在1,25-(OH)2D3限定培养中表达NGF

doi:国家自然科学基金资助课题

脑皮质星形细胞在1,25-(OH)₂D₃限定培养中表达NGF

李贤慧¹，程熠²，刘雅文³，王刚¹，李积胜¹

1.　武警医学院生物化学与分子生物学教研室，天津300162； 2.吉林大学基础医学院生物化学与分子生物学教研室，吉林长春130021；3.吉林大学公共卫生学院流行病学教研室，吉林长春130021

收稿日期:2003-11-25 修回日期:1900-01-01 出版日期:2004-05-28 发布日期:2004-05-28

Expression of nerve growth factor gene in astrocytescultured in 1,25-(OH)₂D₃ defined medium

LI Xian-hui¹, CHENG Yi², LIU Ya-wen³ , WANG Gang¹, LI Ji-sheng¹

1. Department of Biochemistry and Molecular Biology, Wujing Medical College, Tianjin 300162, China;2. Departmentof Biochemistry and Molecular Biology， School of Basic Medical Sciences, Jilin University, Changchun 130021,China;3. Department of Epidemiology, School of Public Health ,Jilin University, Changchun 130021,China

Received:2003-11-25 Revised:1900-01-01 Online:2004-05-28 Published:2004-05-28

摘要/Abstract

摘要： 目的：研究原代中枢神经胶质细胞在1,25-二羟维生素D₃［1,25-(OH)₂D₃］化学限定培养中神经生长因子（NGF）基因转录及表达。方法：用新生乳鼠脑皮质制备星形胶质细胞，采用浓度0、10_-7、10^-8、10^-9、10^-10、10^-11及10^-12 mol•L^-1 1,25-(OH)₂D₃化学限定无血清培养液培养后，RT-PCR法检测NGF mRNA转录,双向夹心-ELISA检测NGF。结果：1,25-(OH)₂D₃化学限定无血清原代培养的神经胶质细胞在1,25-(OH)₂D₃作用下可分泌NGF，实验中1,25-(OH)₂D₃有效浓度范围10^-11~10_-7mol•L^-1；1,25-(OH)₂D₃培养6 h后开始检测到mRNA的表达，12 h时表达最高，36 h后检测不到NGF mRNA的表达；而NGF蛋白的ELISA反应持续时间大于48 h。结论：星形神经胶质细胞的NGF基因可被甾体激素1,25-(OH)₂D₃激活并转录表达。

关键词: 骨化三醇, 神经生长因子

Abstract: Objective To investigate the expression of nerve growth factor (NGF) gene in astrocytes cultured in chemical 1,25-(OH)₂D₃ defined medium. Methods The astrocytes prepared from brain of neonatal rats were cultured in a chemical 1,25-(OH)₂D₃ defined serum-free medium with various doses of 1,25-(OH)₂D₃ （0，10_-7，10^-8-8，10^-9-9，10^-10，10^-11 and 10^-12 mol•L^-1 ）. NGF mRNA transcripts were detected by RNA-based polymerase chain reaction method. Secretory NGF protein quantity were measured with a double-site ELISA with monoclonal anti-NGF antibody. Results NGF synthesis and secretion were induced in astrocytes cultured in chemical 1,25-(OH)₂D₃ defined serum-free medium. 1,25-(OH)₂D₃ were effective in the concentration range of 10_-7 to 10^-11 mol•L^-1. The expression of mRNA was detected 6 hours after cultured in 1,25-(OH)₂D₃ medium,the level of expression of mRNA approached to peak value at 12th hour, and NGF mRNA was not found after 36 hours. The reaction period of ELISA of NGF protein was above 48 hours.Conclusion NGF gene in central nervous system (CNS) can be regulated by 1,25-(OH)₂D₃.

Key words: calcitriol, nerve growth factor

中图分类号:

Q565

李贤慧，程熠，刘雅文，王刚，李积胜. 脑皮质星形细胞在1,25-(OH)₂D₃限定培养中表达NGF[J]. J4, 2004, 30(3): 398-400.

LI Xian-hui, CHENG Yi, LIU Ya-wen , WANG Gang, LI Ji-sheng. Expression of nerve growth factor gene in astrocytescultured in 1,25-(OH)₂D₃ defined medium[J]. J4, 2004, 30(3): 398-400.

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脑皮质星形细胞在1,25-(OH)₂D₃限定培养中表达NGF

Expression of nerve growth factor gene in astrocytescultured in 1,25-(OH)₂D₃ defined medium

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