5-Aza-CdR对HT22小鼠海马神经元细胞中NR2B磷酸化的影响

Effects of 5-Aza-CdR on p-Y1472 NR2B in the HT22 cells

目的:研究5-氮杂-2′-脱氧胞苷对HT22小鼠海马神经元细胞中NR2B 1472位点的酪氨酸磷酸化(p-Y1472 NR2B)的影响。方法:使用小鼠海马神经元细胞系HT22,对照组细胞用0.1%BSA处理,处理组细胞用10μmol/L 5-Aza-CdR处理。采用real time RT-PCR检测NR2B mRNA的表达;Western Blot检测NR2B和磷酸化NR2B-Y1472(p-Y1472 NR2B)的表达水平;MTS法检测细胞活力;流式细胞分析技术检测细胞周期。结果:HT22细胞在10μmol/L 5-Aza-CdR作用下,NR2B-Y1472的磷酸化水平升高,细胞活力下降,细胞周期受到阻滞。结论:5-Aza-CdR对小鼠行为记忆能力的影响可能与NR2B-Y1472磷酸化相关。

Objective:To study the effects of 5-Aza-2'-deoxycytidine on the tyrosine phosphorylation of NR2B 1472 site(p-Y1472 NR2B)in the HT22 cells.Methods:The mice hippocampal neuron cell line HT22 was used in vitro experiment,cells in the control group were treated with 0.1%BSA,and the treatment group were treated with 10μmol/L 5-Aza-CdR.The mRNA level of NR2B were measured by real time RT-PCR.NR2B and p-Y1472 NR2B were measured by Western Blot.Cell activity was detected by MTS method.Cell cycle was detected by flow cytometry.Results:In vitro experiments showed that 10μmol/L 5-Aza-CdR can increased the phosphorylation level of NR2B-Y1472,decreased the cell viability and arrested cell cycle.Conclusions:The effects of 5-Aza-CdR on behavioral memory ability may be related to the phosphorylation NR2B-Y1472.