中国现代医学杂志2017，Vol.27Issue(25)：8-12,5.DOI:10.3969/j.issn.1005-8982.2017.25.002
大鼠心肌细胞H9C2活性氧水平检测方法探究
Investigation of detection method of ROS in H9C2 cells
丁雨 1陈雪梅 1吴思思1
作者信息
- 1. 四川大学华西医院公共实验技术中心,四川成都610041
- 折叠
摘要
Abstract
Objective To compare three different methods for ROS measurement,and analyze their advantages and shortcomings so as to select a suitable and accurate one for detecting ROS in hypoxia-reoxygenation (H-R)model of rat myocardial cell line H9C2.Methods H9C2 cells were plated in an appropriate confluence (80％-90％).The experimental group (H-R 6 h) was exposed to hypoxia for 6 h followed by reoxygenation for 2 h,the control group (CON) was cultured in normoxic condition.Then DCFH-DA fluorescent probe was added into H9C2 cells.After incubation,the fluorescent intensity was detected by fluorescent microscopy,flow cytometer and fluorescence multiscan.Then these data were analyzed with appropriate analysis software.Results Compared with the control group,the results of flow cytometer [CON (152,690 ± 34,104) FU/μg,H-R 6 h (325,669 ± 12,755) FU/μg] or fluorescence multiscan [CON (282.3 ± 12.57) FU/μg,H-R 6 h (1,274 ± 37.05) FU/μg] showed that the ROS level of the experimental group was significantly higher than that of the control group (P ＜ 0.05),but the results from fluorescence microscopy could not distinguish the experimental group from the control group.Conclusions Fluorescence multiscan can better determine the ROS level in H9C2 cells.关键词
缺氧/复氧/H9C2细胞/活性氧Key words
hypoxia-reoxygenation/H9C2 cell/ROS分类
医药卫生引用本文复制引用
丁雨,陈雪梅,吴思思..大鼠心肌细胞H9C2活性氧水平检测方法探究[J].中国现代医学杂志,2017,27(25):8-12,5.
