Sa Zhe-Yan, Guo Shi-Yu, Shan Li-Dong, Gong Shan, Gao Hong, Hisamitsu Tadashi, Jiang Xing-Hong
Department of Neurobiology and Medical Psychology, Laboratory of Aging and Nervous Diseases, Medical College, Suzhou University, Suzhou 215123, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2010 Aug;26(3):359-64.
To observe the effect of nitric oxide (NO) on the differentiation of neural stem cells (NSCs) derived from subventricular zone (SVZ) of neonatal rats in vitro.
Conventional method was used to isolate and culture the NSCs from SVZ. Diethylenetriamine/NO(DETA/NO) was used as NO donor and Nitro-L-arginine methylester (L-NAME) was used as inhibitor of nitric oxide synthase (NOS). The immunofluorescence was used to identify the expression of nestin (a marker of NSCs), beta-III-tubulin (Tuj-1, a marker of neurons), glial fibrillary acidic protein (GFAP, a marker of astrocytes) and nNOS. The concentration of NO in medium was measured by Greiss assay.
Cultured neurospheres were nestin-, BrdU- and nNOS-positive. After treatment with 40 micromol/L, 50 micromol/L and 60 micromol/L of DETA/NO for 5 days, the concentration of NO released was increased significantly (P < 0.01) as compared with that of the control group. The percentage of both differentiated neurons and astrocytes was increased significantly (P < 0.01 and P < 0.05) as compared with that of the control group. After treatment with 100 micromol/L, 150 micromol/L and 200 micromol/L of L-NAME for 5 days, the concentration of NO released was decreased as compared with that of the control group (P < 0.05). The percentage of both differentiated neurons and astrocytes were decreased as compared with that of the control group (P < 0.05).
NO could directly promote the differentiation of NSCs derived from rat subventricular zone in vitro.
观察一氧化氮（NO）对新生大鼠脑室下区（SVZ）神经干细胞（NSCs）体外分化的影响。
采用常规方法从SVZ分离培养NSCs。以二乙三胺/NO（DETA/NO）作为NO供体，硝基-L-精氨酸甲酯（L-NAME）作为一氧化氮合酶（NOS）抑制剂。运用免疫荧光法鉴定巢蛋白（NSCs的标志物）、β-III微管蛋白（Tuj-1，神经元的标志物）、胶质纤维酸性蛋白（GFAP，星形胶质细胞的标志物）和nNOS的表达。采用格里斯试剂法测定培养基中NO的浓度。
培养的神经球呈巢蛋白、BrdU和nNOS阳性。用40μmol/L、50μmol/L和60μmol/L的DETA/NO处理5天后，与对照组相比，释放的NO浓度显著升高（P<0.01）。分化的神经元和星形胶质细胞的百分比与对照组相比均显著增加（P<0.01和P<0.05）。用100μmol/L、150μmol/L和200μmol/L的L-NAME处理5天后，与对照组相比，释放的NO浓度降低（P<0.05）。分化的神经元和星形胶质细胞的百分比与对照组相比均降低（P<0.05）。
NO可直接促进体外培养的大鼠脑室下区神经干细胞的分化。
