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目的 研究未折叠蛋白反应 (UPR)在神经分化过程中的作用。 方法 应用全反式维甲酸 (RA)诱导鼠胚胎干细胞 (ES) ,通过免疫细胞化学和RT PCR方法检测神经组织特异性标志物的表达 ,建立以RA诱导ES细胞得到的神经分化的初步模型 ,再分别用RT PCR和Westernblot方法检测模型中UPR分子的表达。 结果 RA诱导后得到大量表达神经特异性标志物的细胞。UPR关键分子Irelα的表达在RA处理组和未经RA处理组中均下降 ,但未经RA处理组中Irelα的表达始终低于同一时间RA处理组细胞 ;Grp78的表达在RA处理组随诱导时间延长逐渐上升 ,但在未经RA处理组Grp 78的表达未见上升。Irelα和Grp78的这种表达变化与RA诱导细胞中神经特异性标志物的表达情况相关。 结论 以RA诱导ES细胞得到表达神经特异性标志物的细胞分化系统 ,可作为研究神经发育的初步模型 ,其模型中UPR分子的表达变化提示 ,UPR通路可能在神经发育过程中起一定作用
Objective To investigate the role of unfolded protein response (UPR) in neural differentiation. Methods The rat embryonic stem cells (ESs) were induced by all-trans retinoic acid (RA). The expression of specific neuronal markers was detected by immunocytochemistry and RT-PCR. A preliminary model of neuronal differentiation induced by RA was established. The expression of UPR in the model was detected by RT PCR and Western blot respectively. As a result, a large number of cells expressing neural specific markers were obtained after RA induction. The expression of Irelα, a key molecule of UPR, was decreased in both RA and non-RA treated groups, but the expression of Irelα in RA-treated group was always lower than that in RA treated group at the same time. The expression of Grp78 increased with the induction time However, the expression of Grp78 did not increase in RA group. This change in expression of Irel [alpha] and Grp78 correlates with the expression of neural-specific markers in RA-induced cells. Conclusion The cell differentiation system expressing neural-specific markers in RA-induced ES cells can be used as a preliminary model for studying neural development. The expression changes of UPR in its model suggest that UPR pathway may play a role in neural development