低氧预适应对神经细胞NG108-15 hamartin基因DNA甲基化的影响  

Effects of hypoxia preconditioning on the hamartin DNA methylation in NG108-15 cell line

作　　者：朱宏伟[1] 田小莉 齐瑞芳 杨洁[2,3] 侯林 时静华 邵国 

机构地区：[1]内蒙古科技大学包头医学院第二附属医院内分泌科,内蒙古包头014030 [2]内蒙古低氧转化医学重点实验室 [3]北京市低氧适应转化医学重点实验室,100053 [4]内蒙古科技大学包头医学院

出　　处：《包头医学院学报》2018年第1期87-88,98,共3页Journal of Baotou Medical College

基　　金：国家自然科学基金(No:81160244;81360316;81460283;81660307);内蒙古自治区高等学校青年科技英才支持计划资助(No:NJYT-13-A10);内蒙古自治区高等学校科研项目(NJZY 207);包头医学院基金(BYJJ-DF201602;BYJJ-YF201615;BSJJ201617;BYJJ-QM201633;BYJJ-QM201656;BYJJ201502);包头市科技计划项目(CX2017-54)

摘　　要：目的:以神经细胞NG108-15为实验材料探讨低氧预适应对神经细胞hamartin基因DNA甲基化的影响。方法:培养神经细胞NG108-15,将其分为空白对照组(C组,不低氧)、低氧对照组(H组,单纯低氧)和低氧预适应组(HP组,给予低氧预适应)3个组。H组细胞暴露于1%的低氧环境中8 h,然后复氧(21%氧气环境)8 h;HP组细胞暴露于1%低氧环境中30 min,复氧(21%氧气环境)30 min,4个循环后暴露于1%的低氧环境中8 h,然后复氧(21%氧气环境)8 h。倒置相差显微镜观察细胞形态,MTT检测细胞存活率,DNA甲基化测序分析hamartin基因DNA甲基化的情况。结果:低氧预适应降低低氧对NG108-15细胞形态的影响,并且改变了hamartin基因promoter区的DNA甲基化情况。结论:低氧预适应神经保护作用可能涉及hamartin基因DNA甲基化变化。Objective：To explore the effect of Hypoxia preconditioning on the hamartin promoter DNA methylation level using NG108-15 as the experimental material.Methods：NG108-15 cell line was cultured and divided into three groups.The control group（C group） was cultured under normal condition;hypoxia group（H group） was exposed to 1% O_2 for 8 h and then underwent 8 h of reoxygenation（21% O_2）;hypoxia precondition group（HP group） was exposed to 1% O_2 for 30 min and then 21 % O_2 for 30 min.This process was repeated 4 times and exposed to 1% O_2 for 8 h and then underwent 8 h of reoxygenation（21% O_2）.The inverted phase contrast microscope was used to observe the morphology of NG108-15 cell,MTT was used to measure the cell viability,and DNA methylation bisulphite sequencing（DMBS） was adopted to measure the hamartin promoter DNA methylation.Results：Hypoxia preconditioning can decrease the effect of hypoxia on the morphology of NG108-15,and at the same time change the hamartin promoter DNA methylation level.Conclusions：The neuroprotective effect of hypoxia preconditioning may involve in the change of hamartin promoter DNA methylation.

关 键 词：低氧预适应 DNA甲基化 HAMARTIN 

分 类 号：R346[医药卫生—基础医学]