N-硝基-L-精氨酸对人胚胎神经干细胞增殖的作用  

Effects of L-NNA on proliferation of neural stem cells from human fetus

作　　者：赵璇[1] 郑慧媛[1] 赵凌宇[2] 吴志新[1] 崔媛媛[1] 王兰[1] 

机构地区：[1]西安医学院组织学与胚胎学教研室,西安710021 [2]西安交通大学医学院

出　　处：《陕西医学杂志》2011年第12期1581-1583,共3页Shaanxi Medical Journal

摘　　要：目的:探讨一氧化氮合酶(NOS)抑制剂N-硝基-L-精氨酸(L-NNA)对人胚胎神经干细胞的增殖作用。方法:取4月龄(120±15d)正常孕妇引产胎儿的大脑皮质组织分离细胞,用无血清的条件培养基,培养原代神经干细胞并用间接免疫荧光化学法进行鉴定。第5天时,在培养基中加入不同浓度的L-NNA,继续培养24h后,用Griess还原法检测培养液中一氧化氮含量;用四甲基偶氮唑蓝比色法(MTT)测定细胞活力。结果:加入不同浓度L-NNA24h后,培养液中一氧化氮量随L-NNA浓度的增高而降低,且显著低于对照组(P<0.05)。MTT法显示经L-NNA作用后的神经干细胞活细胞数较对照组明显增加(P<0.05),并呈浓度相关性。结论:L-NNA对培养状态下的人胚胎神经干细胞增殖有促进作用。Objective： To study the effects of N-nitro-L-arginine（L-NNA）,one of selective nitric oxide synthase inhibitors,on the proliferation of human fetal neural stem cells（NSCs）.Methods： The primary NSCs from the cerebral cortex of 4-month-old human fetuses were cultivated in the serum-free medium,and identified with indirect immunofluorescence cytochemistry.On day 5,the NSCs were cultured in the medium supplemented with different concentrations of L-NNA,and 24h later the NO content in the medium wasmeasured with the Griess assay.Then half of the cultured cells were measured with MTT assay to determine the amount of living cells.Results： The NO content in the experimental groups was decreased with the increasing of L-NNA concentration,and significantly higher than that in the control group.The amount of living cells in the experimental groups was significantly increased in a concentration-dependent manner compared with that in the control group.Conclusion： L-NNA can promote the proliferation of NSCs from human fetus in vitro.

关 键 词：干细胞/药物作用 硝基精氨酸/拮抗剂和抑制剂 细胞培养 细胞计数 荧光抗体技术 间接 

分 类 号：R421[医药卫生—临床医学]