大豆异黄酮对大鼠海马神经干细胞分化的影响

Effect of Soy bean Isoflavones on Dysdifferentiation of Cultured Hippocampus Neural Stem Cells Originated from Neonatal Rat

目的:研究过氧亚硝基阴离子(ONOO-)对新生大鼠海马神经干细胞(NSCs)分化的影响以及大豆异黄酮(ISO)对抗ONOO-致脑神经细胞氧化损伤的作用及其可能的作用机制。方法:制备新生大鼠海马神经干细胞并经免疫荧光鉴定。以吗啉-斯得酮亚胺(SIN-1,1 mmol.L-1)作为ONOO-供体引起海马神经干细胞出现分化障碍,再分别用氧合血红蛋白(HbO2,0.05 mmol.L-1),超氧化物歧化酶/过氧化氢酶(SOD/catalase,50 U.mL-1),褪黑激素(0.5 mmol.L-1)以及ISO(0.1mg.L-1)进行干预,免疫荧光检测神经干细胞及其分化情况。结果:SIN-1引起海马神经干细胞巢蛋白阳性(NSE+)细胞数量明显减少,已分化神经细胞形态变化很明显。单独使用一氧化氮阴离子(NO.)清除剂HbO2或超氧阴离子(O 2-.)清除剂SOD/catalase可以增加NSE+细胞数量,改善形态变化。但ONOO-清除剂(褪黑激素)和ISO能够更明显地增加NSE+细胞数量并改善形态变化,尤以前者效果更加明显。结论:ONOO-可对新生大鼠海马神经干细胞的分化产生不利影响。ISO可以对抗ONOO-引起的海马神经干细胞的分化障碍,在脑神经细胞氧化损伤的修复方面具有良好的应用潜力。

Objective：To study the effects of peroxynitrite anoin（ONOO-） on the differentiation of hippocampus neural stem cells（NSCs） of primary rat,and the action and potential mechanisms of soy bean isoflavones against oxidative damage produced by ONOO-in brain nerve cells.Method： Hippocampus NSCs of primary rat were prepared according to the methods of trypsin digestion combined with mechanical dissociation,and differentiated nerve cells were identified with the method of immunofluorescence.Dysdifferentiation of hippocampus NSCs were induced by SIN-1（1 mmol·L-1） as a ONOO-donor,then the cells were treated with HbO2（0.05 mmol·L-1）,SOD/catalase（50 U·mL-1）,melatonin（0.5 mmol·L-1） and isoflavones（0.1 mg·L-1）,respectively,and the differentiation of NSCs was detected with the method of immunofluorescence.Result： Differentiation of hippocampus NSCs were limited by the SIN-1,the number of nestin-positive（NSE＋） cells was significantly decreased and the morphological changes of differentiated neural cells were obvious.The number of NSE＋ cells was increased and the morphological changes were alleviated with the treatment of NO-scavenger（HbO2）or O-2·scavenger（SOD/catalase）alonely,but that could be alleviated significantly with the treatment of ONOO-scavenger（melatonin） or soy bean isoflavones and the former was better.Conclusion： The dysfunction in the differentiation of hippocampus NSCs of neonatal rat can be produced by ONOO-,which can be improved moderately by soy isoflavones showing the potential of isoflavones in application to the repair of oxidative damages in brain nerve cells.