硝普钠对小鼠神经干细胞增殖的抑制作用

Inhibitive effects of sodium nitroprusside on the proliferation of neural stem cells from mouse

目的 探讨外源性一氧化氮对小鼠神经干细胞增殖的抑制作用。方法 用含生长因子的培养基 ,原代培养神经干细胞。第 5d时 ,取部分原代培养细胞于培养基中加入不同浓度的硝普钠进行培养。 2 4h后用Griess还原法检测细胞上清液中一氧化氮的释放量。将这些细胞中的一半进行四甲基偶氮唑蓝比色法 (MTT法 )试验 ,另一半细胞换成无硝普钠的条件培养基继续培养 2 4h ,再行MTT试验。另取原代培养细胞经硝普钠作用 2 4h后 ,行血清诱导分化试验。结果 2 4h后细胞上清液中一氧化氮释放量随硝普钠浓度的增高而增加 ;MTT试验反映经硝普钠作用后的神经干细胞活细胞数较对照组明显减少 ,其减少的程度与硝普钠浓度呈正相关 ;经硝普钠作用后的神经干细胞 ,在去除硝普钠后仍可保持旺盛的增殖能力并能被血清诱导分化为神经细胞样细胞。

Objective To explore the inhibitive effect of ectogenetic nitric oxide(NO) on the proliferation of neural stem cells(NSCs) from mouse. Methods The primary culture NSCs were cultivated in medium containing EGF. After five days, some of this primary culture cells were cultivated in medium supplemented with different concentration of sodium nitroprusside (SNP). The release of NO from SNP was measured by the Griess assay in the medium after 24 hours; Half of the cells cultured with SNP were measured by MTT assay to reflect the amount of living cells in it, and the remaining cells were kept on culturing in medium without SNP for another 24 hours, and then it was measured by MTT assay. Some of the primary culture cells were cultivated with SNP for 24 hours, and then induced to differentiate by serum. Results The release of NO was increasing along with the enhancement of the concentration of SNP; The amount of living cells reflected by MTT test in experiment groups was less compared with that in the control group and the level of decrease was correlated positively with the concentration of SNP; The NSCs still had the ability to proliferate and could be induced to differentiate into neuron and glial cell in medium when SNP had been removed. Conclusion Ectogenetic nitric oxide can inhibit the proliferation of NSC from mouse in vitro.