神经生长因子对小鼠海马神经元细胞系HT22分化及DNMT表达的影响  

Involvement of nerve growth factor in mouse hippocampal neuronal cell line(HT22)differentiation and underlying role of DNA methyltransferases

作　　者：姜树原[1] 王强[2] 张晓璐 刘晓蕾[1] 谢雅彬 巴德仁贵 刘友[2] JIANG Shu-Yuan;WANG Qiang;ZHANG Xiao-Lu(Biomedical Research Center,Institute of Basic Medicine and Forensic Medicine,Baotou 014040,Inner Mongolia,China)

机构地区：[1]包头医学院生物医学研究中心,内蒙古自治区低氧转化医学重点实验室,内蒙古包头014040 [2]包头医学院基础医学与法医学院

出　　处：《中国老年学杂志》2020年第10期2144-2147,共4页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(81660307);内蒙古自然科学基金项目(2019MS08052,2016MS(LH)0307);内蒙古自治区高等学校科研项目(NJZY17251);包头医学院科学研究基金项目(BYJJ-YF201615,BYJJ-YF201719)。

摘　　要：目的探讨神经生长因子(NGF)对小鼠海马神经元细胞系HT22分化及DNMT表达的影响。方法小鼠海马神经元细胞系HT22分为未分化组(不经特殊处理)和分化组(50 ng/ml NGF-β处理细胞)。通过活细胞动态成像与分析系统IncuCyte分析NGF是否参与小鼠海马神经元细胞系HT22的分化。实时定量聚合酶链反应和Western印迹法检测DNA甲基转移酶(DNMT)基因和蛋白质表达。应用甲基转移酶活性试剂盒检测DNMTs活性。应用流式细胞术检测细胞周期。结果NGF处理后,分化组与未分化组HT22细胞相比DNMT1、DNMT3A、DNMT3B的mRNA表达水平显著升高了13倍、8倍、5倍(P<0.05),其中DNMT1、DNMT3A蛋白质表达量显著增加1.5倍(P<0.05),DNMTs活性增强(P<0.05),S期细胞百分比显著增加。结论NGF诱导的HT22细胞的分化与DNMTs的激活有关。Objective To investigate the influence of nerve growth factor(NGF)on HT22 cell differentiation and DNMT expression.Methods The mouse hippocampal neuron cell line HT22 was divided into undifferentiated group(without special treatment)and differentiated group(50 ng/ml NGF-βtreated cells).IncuCyte was used to determine whether NGF was involved in differentiation of HT22.Quantitative PCR and western blot were used to measure gene and protein expression of DNMT as well as the activity of DNMTs.Results Treatment with NGF was found to upregulate both gene and protein expressions as well as total activity of DNMTs in differentiating HT22 cells.Compared to undifferentiating cells,the percentage of differentiating cells at S phase increased significantly when incubated with NGF.In undifferentiated cells,NGF failed to induce gene and protein expressions and activity of DNMTs.Conclusions Differentiation of HT22 cells induced by NGF involves the activation of DNMTs pathway.

关 键 词：DNA甲基转移酶 HT22细胞系 神经生长因子 

分 类 号：R338[医药卫生—人体生理学]