解剖学报

解剖学报 ›› 2013, Vol. 44 ›› Issue (5 ): 635-640.doi: 10.3969/j.issn.0529-1356.2013.05.010

• 神经生物学 • 上一篇    下一篇

一氧化氮对星形胶质细胞轴突生长因子-1表达及迁移的影响

施月 张晔 邵杰 姚扬明 夏春林*   

  1. 苏州大学医学部博习临床研究所暨细胞神经生物研究室，苏州 215123
  • 收稿日期:2012-07-09 修回日期:2012-09-20 出版日期:2013-10-06 发布日期:2013-10-06
  • 通讯作者: 夏春林 E-mail:xclcnb@suda.edu.cn；clxiasz@gmail.com

Effects of nitric oxide on netrin-1 expression and migration of astrocytes

SHI Yue  ZHANG Ye  SHAO Jie  YAO Yang-ming  XIA Chun-lin*   

  1. Boxi Institute of Clinical Anatomy and Cytoneurobiology Laboratory, Medical College of Soochow University, Suzhou 215123, China
  • Received:2012-07-09 Revised:2012-09-20 Online:2013-10-06 Published:2013-10-06

摘要：

目的 探讨一氧化氮(NO)对星形胶质细胞中轴突生长因子-1(netri-1)的表达变化以及对细胞迁移的影响。方法 采用硝普钠（SNP）作为NO供体处理星形胶质细胞，通过振荡培养和差速贴壁法分离纯化新生SD大鼠星形胶质细胞，接种至培养板，分为实验组和对照组，每组6个样本，实验组用50μmol/L SNP处理星形胶质细胞，通过划痕法观察细胞的迁移，并采用Western blotting 和免疫细胞化学方法分别检测处理前后的星形胶质细胞中netrin-1蛋白的表达变化。 结果 SNP处理后，实验组的星形胶质细胞与对照组相比，划痕区细胞明显增多，星形胶质细胞netrin-1表达随着时间的推移出现先升高后降低趋势，于48 h达到峰值 (P<0.01)。 结论 SNP使星形胶质细胞netrin-1表达水平发生变化和影响星形胶质细胞迁移，提示netrin-1可能通过NO的调控而影响星形胶质细胞迁移。

关键词: 星形胶质细胞, 轴突生长因子-1, 一氧化氮, 细胞迁移, 免疫印迹法, 大鼠

Abstract:

Objective To investigate effects of NO on netrin-1 expression and migration of astrocytes. Methods Astrocytes were treated with a certain concentration of NO donor sodium nitroprusside (SNP) in vitro.The astrocyte migration was observed and the expression of netrin-1 at the different times after treatment was detected. Purified cultures of astrocytes were prepared from neonatal rats, based on the differential properties of developmental time-course and cellular adhesions. Astrocytes were inoculated to culture plates and were divided into experimental and control groups (n=6).The experimental groups were treated with 50μmol/L NO donor SNP. Cell migration was observed by scratching. The expression of netrin-1 after treatment was examined by Western blotting and immunocytochemistry. Results The speed of astrocyte migration increased after SNP treatment. The expression of netrin-1 in the protein level began to increase at 6 hours and reached the peak at 48 hours, compared with the control group (P<0.01), and then declined. Conclusion SNP may increase the expression of netrin-1 protein and promote astrocytes migration, indicating that netrin-1 promoting astrocytes migration maybe controled by NO.

Key words: Astrocyte, Netrin-1, Nitric oxide, Cell migration, Western blotting, Rat