摘要
目的:观察全反式维甲酸(all-trans retinoic acid,ATRA)体外对大鼠中脑神经干细胞(neural stem cells,NSCs)的诱导分化作用及其可能的分子机制。方法:分离培养孕14～15 d大鼠中脑NSCs,使用1.0μmol/L ATRA诱导1、3、5和7 d,以免疫组织化学染色观察ATRA对NSCs分化为多巴胺能神经元的影响;RT-PCR分析维甲酸(retiadic acid,RA)各受体表达情况。结果:ATRA诱导后多巴胺能神经元特异性酪氨酸羟化酶(TH)染色呈阳性,细胞有多个细长突起,与对照组比较,诱导7 d后多巴胺神经元特异性TH染色阳性细胞率均明显升高(P<0.01);各诱导组RA受体βmRNA的表达量均明显升高(P<0.01),而且存在时间依赖性。结论:ATRA能显著提高NSCs分化为多巴胺能神经元的比例,同时发现细胞维甲酸A受体(RAR)βmRNA表达增加,RARβ激活可能促进NSCs向多巴胺神经元方向的分化。
Objective： To observe the role of all-trans retinoic acid（ATRA） in inducing differentiation of rat midbrain-derived neural stem cells（NSCs） and its possible molecclar mechanism.Methods： Pregnant 14-15 day rats midbrain-derived NSCs were obtained by mechanical isolation and serum-free culture.Neural stem cells were induced by 1μmol/L ATRA on day 1,day 3,day 5 and day 7.The roles of ATRA in inducing the differentiation of NSCs were observed with immunofluorescent staining,and the expression of retinoic acid receptor（RAR） gene was determined with semi-quantitative RT-PCR.Results： After ATRA induction,neural stem cells were positive for tyrosine hydroxylase（TH）,the rate of TH-positive cells significantly increased in the ATRA 7-day groups（P 0.01）.Compared with the control group,RARβmRNA expression significantly increased in each group（P 0.01）.That ATRA could induce the expression of RARβmRNA was a time dependent manner.Conclusion： ATRA can greatly increase the percentage of neurons differentiation from rat embry-onic neural stem cells and up-regulate the expression of RARβmRNA in rat midbrain-derived NSCs.Activation of RARβ may promote the differentiation of NSCs into dopaminergic neurons.
出处
《温州医学院学报》
CAS
2011年第1期6-9,12,共5页
Journal of Wenzhou Medical College
基金
浙江省科技厅科研基金资助项目(2007C23026)
浙江省卫生厅科研基金资助项目(2007B146)
温州市科技局科研基金资助项目(Y20070042)
关键词
全反式维甲酸
细胞分化
神经干细胞
多巴胺神经元
大鼠
all-trans retinoic acid
cell differentiation
neural stem cells
dopaminergic neurons
rats
