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目的探讨外源性一氧化氮(NO)对培养状态下的新生大鼠神经干细胞/前体细胞(NSCs/NPs)分化的影响。方法采用常规方法分离新生大鼠脑室下带(SVZ)组织,进行NSCs/NPs体外培养。用二乙烯三胺/一氧化氮聚合物(DETA/NO)作为外源性NO供体培养NSCs/NPs。用免疫细胞化学方法检测NSCs/NPs标记巢蛋白(nestin)、神经元标记神经元特异性微管相关蛋白(Tuj-1)和星型胶质细胞标记胶质原纤维酸性蛋白(GFAP)。同时用Greiss还原法检测培养液中总NO的浓度。结果原代或次代培养的神经球均为nestin阳性;DETA/NO对体外培养的NSCs/NPs作用5d后,40μmol/L、50μmol/L、60μmol/L DETA/NO实验组分别释放出(62.0±6.4)μmol/L、(64.8±15.7)μmol/L、(68.5±11.6)μmol/L的NO,相应实验组分化的神经元数分别为(53.1±4.7)%、(54.1±6.9)%、(63.8±9.5)%,较对照组[(38.8±7.2)%],显著增加,差异有高度统计学意义(P<0.01);50μmol/L、60μmol/LDETA/NO实验组分化的星型胶质细胞数分别为(38.2±6.7)%、(41±10.5)%,较对照组[(32.8±5.5)%],有所增加,差异有统计学意义(P<0.05)。结论外源性NO主要促进体外培养的NSCs/NPs向神经元方向分化。
Objective To investigate the effects of exogenous nitric oxide (NO) on the differentiation of neural stem / progenitor cells (NSCs / NPs) in cultured neonatal rats. Methods SVZ tissues of neonatal rats were isolated by routine method and cultured in vitro. NSCs / NPs were cultured with diethylenetriamine / nitric oxide polymer (DETA / NO) as exogenous NO donor. NSCs / NPs labeled nestin, neuron-specific neuronal specific microtubule-associated protein (Tuj-1) and astroglial marker glial fibrillary acidic protein (GFAP) were detected by immunocytochemistry. Meanwhile, the concentration of total NO in culture solution was detected by Greiss reduction method. RESULTS: After treated with DETA / NO for 5 days, the neurons in 40μmol / L, 50μmol / L and 60μmol / L DETA / NO groups released (62.0 ± (53.1 ± 4.7)% and (54.1 ± 6.9)% of the neurons in the corresponding experimental groups were significantly higher than those in the control group (P <0.05) (63.8 ± 9.5)%, which was significantly higher than that in the control group [(38.8 ± 7.2)%], the difference was highly statistically significant (P <0.01) (38.2 ± 6.7)% and (41 ± 10.5)%, respectively, which was significantly higher than that of the control group [(32.8 ± 5.5)%], the difference was statistically significant (P <0.05). Conclusion Exogenous NO mainly promotes the differentiation of NSCs / NPs into neurons in vitro.