人嗅黏膜神经干细胞体外培养及氟西汀对其增殖和神经发生的影响  

Culture of Human Olfactory Mucosa Neural Stem Cells In Vitro and Effects of Fluoxetine on Their Proliferation and Neurogenesis

作　　者：谢扬智 李乾露 杨德雨[1,2] 房亮[2] 廖娟[2] 李远[2] 王娟[1] 刘莉[3] 江洪[4] 李志伟[1] Xie Yangzhi Li Qianlu Yang Deyu Fang Liang Liao Juan Li Yuan Wang Juan Liu Li Jiang Hong Li Zhiwei(Department of Neurology, Yongchuan Hospital Chongqing Medical University, Chongqing 402160, China ZCentral Laboratory, Yongchuan Hospital, Chongqing Medical University, Chongqing 402160, China Department of Health Management, Yongchuan Hospital Chongqing Medical University, Chongqing 402160, China Department of Otolaryngolory, Yongchuan Hospital Chongqing Medical University, Chongqing 402160, China)

机构地区：[1]重庆医科大学附属永川医院神经内科,重庆402160 [2]重庆医科大学附属永川医院中心实验室,重庆402160 [3]重庆医科大学附属永川医院体检中心,重庆402160 [4]重庆医科大学附属永川医院耳鼻喉颈科,重庆402160

出　　处：《中国细胞生物学学报》2016年第12期1480-1486,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:31300917);重庆市自然科学基金计划(批准号:cstc2016jcyj A0452)资助的课题~~

摘　　要：人嗅黏膜神经干细胞是新近发现的可用于神经精神疾病研究的良好材料,同时,抗抑郁治疗促进脑部神经干细胞增殖及向神经细胞转化。然而,嗅黏膜神经干细胞体外培养方法及抗抑郁药对其增殖及神经发生的影响尚不清楚。该研究采用组织块培养获取嗅细胞,神经球培养法纯化神经干细胞,Nestin免疫荧光验证神经干细胞特异性。加入促神经分化培养基诱导神经干细胞向神经细胞分化,MAP2(microtubule associated protein 2)、TUJ1(β3-tubulin)免疫荧光验证神经细胞特异性。采用CCK-8和蛋白质分子印记技术分别检测抗抑郁药对嗅黏膜神经干细胞增殖及分化的影响。结果显示,体外培养嗅黏膜神经干细胞呈球状聚集;Nestin、MAP2、TUJ1免疫荧光阳性;抗抑郁药促进嗅黏膜神经干细胞增殖、促进其向神经细胞转化,TUJ1表达上升。综上所述,抗郁药能够促进神经干细胞增殖及神经发生,可能为其抗抑郁治疗的机制之一。Human olfactory neural stem cells are satisfactory materials apllied in neuropsychiatry diseases recently discovered. Antidepressants could promote proliferation and neurogenesis of neural stem cells in human. However, culture method in vitro and effects of antidepressants on their proliferation and neurogenesis remains unknown. In the present study, tissue piece culture method and neurosphere method were used to harvest olfactory cells and purify neural stem cells separately. Nestin immunofluorescence was performed to verify the specificity of neural stem cells. Neural basal medium was used to induce the differentiation into neuron from neural stem cells. TUJ1 and MAP2 immunofluorescence were performed to verify the specificity of neuron. CCK-8 and Western blot were applied to measure effects of antidepressants on proliferation and differentiation of neural stem cells. Results showed that olfactory neural stem cells aggregated as neurosphere. Immunofluoscence assay showed Nestin, TUJ1, MAP2 positive in the treated cells. Compared with control group, antidepressants enhance the proliferation of neural stem cells, also, the expression of TUJ1 were up-regulated. Antidepressants could increase neurogenesis of neural stem cells which was likely to be one of the mechanisms in antidepressant treatment.

关 键 词：嗅黏膜 神经干细胞 神经发生 抗抑郁药 组织块培养 

分 类 号：R749.4[医药卫生—神经病学与精神病学]