欢迎访问《解剖学报》官方网站！今天是
English
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
Effects of nitric oxide on netrin-1 expression and migration of astrocytes
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。
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.
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
Astrocyte / Netrin-1 / Nitric oxide / Cell migration / Western blotting / Rat
［1］Thomassen MJ, Buhrow LT, Connors MJ, et al. Nitric oxide inhibits inflammatory cytokine production by human alveolar macrophages［J］. Am J Respir Cell Mol Biol, 1997, 17 (3) :279-283.
［2］Chen A, Kumar SM, Sahley CL, et al. Nitric oxide influences injury-induced microglial migration and accumulation in the leech CNS［J］. J Neurosci, 2000, 20 (3) :1036-1043.
［3］Knipp S, Bicker G. Regulation of enteric neuron migration by the gaseous messenger molecu- les CO and NO［J］. Development, 2009, 136 (1) : 85-93.
［4］Li ChP, Zhang Y, Xia ChL,et al.The expression of nestin and SSEA-1 in the rat type-2 astrocytes ［J］.Acta Anatomica Sinica, 2007, 38(2): 153-157. (in Chinese)
李春鹏, 张晔, 夏春林, 等. 巢蛋白和阶段特异性胚胎抗原1在大鼠2型星形胶质细胞中的表达［J］. 解剖学报, 2007, 38(2): 153-157.
［5］Benarroch EE. Neuron-astrocyte interactions partnership for normal function and disease in the central nervous system［J］. Mayo Clin Proc, 2005, 80 (10) : 1326-1338.
［6］Aloisi F, Ria F, Adorini L. Regulation of T-cell responses by CNS antigen-presenting cells: different roles for microglia and astrocytes［J］. Immunol Today, 2000, 21 (3) : 141-147.
［7］Bahr M, Przyrembel C, Bastmeyer M. Astrocytes from adult rat optic nerves are nonpermissive for regenerating retinal ganglion cell axons［J］. Exp Neurol, 1995, 131 (2) : 211-220.
［8］Ischiropoulos H, al-Mehdi AB. Peroxynitrite-mediated oxidative protein modifications［J］. FEBS Lett, 1995, 364 (3) : 279-282.
［9］Benarroch EE. Neuron-astrocyte interactions: partnership for normal function and disease in the central nervous system［J］. Mayo Clin Proc, 2005, 80 (10) : 1326-1338.
［10］Bicker G. Stop and CO with NO: nitric oxide as a regulator of cell motility in simple brains［J］. Bioessays, 2005, 27 (5) : 495-505.
［11］Rajasekharan S, Kennedy TE. The netrin protein family［J］. Genome Biology, 2009, 10 (9): 239.
［12］Jarjour AA, Manitt C, Moore SW, et al. Netrin-1 is a chemorepellent for oligodendrocyte precursor cells in the embryonic spinal cord［J］. Neuroscience, 2003, 23 (9) :3735-3744.
［13］Manitt C, Colicos MA, Thompson KM, et al. Widespread expression of netrin-1 by neurons and oligodendrocytes in the Gadult mammalian spinal cord［J］. Neuroscience, 2001, 21 (11): 3911-3922.
［14］Tessier-Lavigne M, Goodman CS. The molecular biology of axon guidance［J］. Science, 1996, 274 (5290) : 1123-1133.
［15］Liu Y, Stein E, Oliver T, et al. Novel role for netrins in regulating epithelial behaviour during lung branching morphogenesis［J］. Curr Biol, 2004, 14 (10): 897-905.
［16］Barallobre MJ, Pascual M, Del Rio JA, et al. The netrin family of guidance factors: emphasis on netrin-1 signalling ［J］. Brain Res Rev, 2005, 49 (1) : 22-47.
/
| 〈 |
|
〉 |
