目的:探讨全反式维甲酸体外促神经干细胞分化为神经元过程中DNA甲基转移酶1和乙酰化组蛋白H3的表达变化。方法:0.5、1.0和4μmol/L全反式维甲酸诱导体外培养的鼠胚神经干细胞,免疫荧光检测NeuN和DAPI,计算各组NeuN/DAPI的比值,免疫印迹检测DNA甲基转移酶1以及乙酰化组蛋白H3表达水平。结果:免疫荧光结果显示给药组NeuN/DAPI比值显著高于对照组。药物浓度在1和4μmol/L时,DNA甲基转移酶1表达水平与对照组相比均显著降低,且有剂量依赖性;而在4μmol/L时,组蛋白H3乙酰化水平较对照组有显著增强。结论:全反式维甲酸促进神经干细胞向神经元的分化可能与DNA甲基转移酶1的表达抑制和组蛋白乙酰化水平增加有关。
Objective: To investigate the expression of DNA methyltransferase 1 (DNMT1) and acetyl-H3 in differentiation from rat neural stem cells (NSC;s) into neurons in vitro induced by all--trans retinoic acid (ATRA). Methods: 0. 5 μmol/L, 1.0 μmol/L, 4 μmol/L ATRA were employed to induce the differentiation of NSCs. Double immunofluorescence labeling for neuronal nuclei (NeuN) and 4', 6-diamidina-2-phenylindole (DAPI), the ratio of NeuN/DAPI was counted. Western blotting was used to detect the level of DNMT1 and acetyl-H3. Results: Immunofluorescence showed that the ratio of NeuN/DAPI in the administration group was significantly higher than that of the control group. At the concentration of 1.0 μmol/L, 4 μmol/L, the levels of DNMT1 in the drug administration group decreased compared to the control group. At the concentration of 4 tanol/L, there was a significant increase in the levels of acetyl-H3 compared with the control group. Conclusion: The effect of ATRA inducing the neuronal differentiation of NSCs could be associated to the inhibition of DNMT1 and enhancement of acetyl-H3.