全反式维甲酸在成年小鼠神经干细胞分化中的调控作用及机制

发布时间：2018-07-21 16:55

【摘要】：目的研究全反式维甲酸(ATRA)对成年小鼠神经干细胞(NSCs)的调控、信号通路及对细胞色素P450(CYP450)家族的影响。方法分离并培养成年小鼠脑室下区NSCs,将细胞分为ATRA组和对照组,ATRA组加入10-6mol/L ATRA,对照组正常培养。采用流式细胞仪检测两组细胞表面标记物,计算神经元、星形胶质细胞、少突胶质细胞和NSCs占总细胞数的百分比;Real-time PCR方法检测CYP450家族相关基因表达情况,筛选出有统计学意义的基因;ELISA法测定细胞色素P450还原酶(CPR)的活性;Western blotting法检测P38 MAPK通路相关蛋白P38、p-P38蛋白的相对表达量。结果 ATRA组NSCs主要是向神经元分化,也有一部分向星形胶质细胞分化,少突胶质细胞较少。与对照组比较,ATRA组P450家族基因中CYP26A1、CYP26B1、CYP26C1表达上调(P均0.05)。ATRA组CPR活性以及p-P38蛋白表达量均较对照组升高(P均0.05)。结论 ATRA促进小鼠NSCs向神经元分化,可能通过上调CYP450中的CYP26家族基因、增加CPR活性以及P38 MAPK通路发挥调控作用。
[Abstract]:Objective to study the regulation, signaling pathway and cytochrome P450 (CYP450) family of adult mouse neural stem cells (NSCs) by all-trans retinoic acid (ATRA). Methods NSCs in the subventricular region of adult mice were isolated and cultured. The cells were divided into ATRA group and control group and 10-6 mol / L ATRA was added into the control group. Cell surface markers were detected by flow cytometry, and the percentage of neurons, astrocytes, oligodendrocytes and NSCs in total cells were calculated. Real-time PCR was used to detect the expression of CYP450 family related genes. The activity of cytochrome P450 reductase (CPR) was detected by Elisa. Western blotting was used to detect the relative expression of P38 MAPK pathway related protein P38 p-P38. Results in ATRA group, NSCs were mainly differentiated into neurons, and some of them were differentiated into astrocytes, with less oligodendrocytes. Compared with the control group, the expression of CYP26A1C1-CYP26B1-CYP26C1 was up-regulated in ATRA group (P 0.05). The CPR activity and p-P38 protein expression in ATRA group were higher than those in control group (P 0.05). Conclusion ATRA can promote the differentiation of NSCs into neurons, which may play a regulatory role by up-regulating CYP26 family gene in CYP450, increasing CPR activity and P38 MAPK pathway.
【作者单位】：苏州大学附属常州第一人民医院;苏州卫生职业技术学院;徐州医科大学;
【基金】：江苏省自然科学基金资助项目(BK20130219) 江苏高校“青蓝工程”科研资助项目
【分类号】：R741

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