摘要
目的:研究氧化应激对人脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells,HUVECs)甲基化和羟甲基化的影响。方法:用1 mmol·L^-1双氧水(H2O2)处理HUVECs诱导细胞内氧化应激,同时以等体积的培养基做为对照。收集处理0 h、1 h、4 h和8 h后的细胞,通过多重反应监测质谱定量(Multiple response monitoring mass spectrometry quantification,MRM-MS)方法检测各组细胞中甲基化率和羟甲基化率的变化。结果:与对照组细胞相比,H2O2处理1 h、4 h和8 h组细胞中DNA甲基化率均显著增加(P<0.05),并随处理时间增加,细胞中DNA甲基化率增加越显著,呈时间依赖性;而1 h、4 h和8 h组细胞中DNA羟甲基化率均显著降低(P<0.05);其中处理1h和4 h后两组之间细胞中DNA羟甲基化率变化不明显,但8 h后,细胞中DNA羟甲基化率显著降低(P<0.05);1h、4 h和8 h组细胞中甲基化率和羟甲基化率总和均显著增加(P<0.05),并且随处理时间增加,细胞中DNA甲基化率和羟甲基化率总和增加越显著,呈时间依赖性。结论:氧化应激增加HUVECs中DNA甲基化率,降低DNA羟甲基化率。
Objective:To investigate the effects of oxidative stress on the methylation and hydroxyl methylation of human umbilical vein endothelial cells(HUVECs).Methods:HUVECs were treated with 1 mmol·L^-1 H2O2 to induce intracellular oxidative stress,equal volume medium were used as control at same time.The cells treated for 0 h,1 h,4 h and 8 h were collected and the changes of methylation rate and hydroxyl methylation rate were detected by multiple response monitoring mass spectrometry(MRM-MS).Results:Compared with the control cells,the DNA methylation rate in the cells treated with H2 O2 for 1 h,4 h and 8 h were significantly increased(P<0.05),and with the increase of treatment time,the DNA methylation rate in the cells were significantly increased,which was time-dependent;While the rate of DNA hydroxyl methylation in the cells treated with H2O2 for 1 h,4 h and 8 h groups were significantly decreased(P<0.05);The rate of DNA hydroxyl methylation between the cells treated with H2 O2 for 1 h with 4 h has no changed,but the rate of DNA hydroxyl methylation in the cells treated with H2O2 for 8 h were significantly decreased(P<0.05);The sum of methylation rate and hydroxyl methylation rate in cells treated with H2O2 for 1 h,4 h and 8 h groups were significantly increased(P<0.05),and with the increase of treatment time,the sum of DNA methylation rate and hydroxyl methylation rate were significantly increased,which was time-dependent.Conclusion:Oxidative stress increased DNA methylation rate and decreases DNA hydroxymethylation rate in HUVECs.
作者
黄菲
陈菲
陈良龙
HUANG Fei;CHEN Fei;CHEN Liang-long(Research center of Gannan Medical University;Gannan Medical University,Ganzhou,Jiangxi 341000)
出处
《赣南医学院学报》
2020年第7期655-658,667,共5页
JOURNAL OF GANNAN MEDICAL UNIVERSITY
基金
江西省教育厅课题(编号:GJJ170882)
赣南医学院科研课题(YB201932)。
关键词
氧化应激
人脐静脉内皮细胞
甲基化
羟甲基化
Oxidative stress
Human umbilical vein endothelial cells
Methylation
Hydroxyl methylation
