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目的探讨一氧化氮(NO)对星形胶质细胞中轴突生长因子-1(netrin-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 the effect of nitric oxide (NO) on the expression of netrin-1 in astrocytes and its effect on cell migration. Methods Astrocytes were treated with nitroglycerin (SNP) as NO donor. The astrocytes were isolated and purified by shaking culture and differential adherent method. The astrocytes were inoculated into the culture plates and divided into experimental group and control group Control group, with 6 samples in each group. In the experimental group, astrocytes were treated with 50μmol / L SNP, the migration of cells was observed by scratch method, and the astrocytes before and after treatment were detected by Western blotting and immunocytochemistry Changes in netrin-1 protein expression in cells. Results Compared with the control group, the numbers of astrocytes in the astrocytes in the experimental group increased significantly after SNP treatment. The expression of netrin-1 in the astrocytes increased first and then decreased with the passage of time. At 48 h peaked (P <0.01). Conclusions SNP can change the expression of netrin-1 in astrocytes and affect astrocyte migration, suggesting that netrin-1 may affect the migration of astrocytes through the regulation of NO.