双波长比例法测定单细胞活性氧探析  

Double wavelengths ratio in detection of the concentrations of reactive oxygen species

作　　者：赵成瑞[1] 崔香丽[2] 吴博威[2] 

机构地区：[1]山西医科大学汾阳学院生理教研室,汾阳032200 [2]山西医科大学生理学系

出　　处：《山西医科大学学报》2011年第1期7-9,共3页Journal of Shanxi Medical University

摘　　要：目的建立双波长比例法测定单细胞活性氧的方法。方法①酶法急性分离成年大鼠心室肌细胞,用离子影像系统测定荧光信号,确定荧光强度值变化相反的两个激发光波长。②BPS-4装置灌流心肌细胞,模拟心肌缺血/再灌注模型及再灌注时应用还原型谷胱甘肽,用双波长比例法测定细胞内活性氧含量变化,验证此法。结果①荧光强度值最大和最小的激发光波长分别是:480 nm和420nm,以F480/F420的比值表示活性氧的含量。②心室肌细胞在模拟缺血期和恢复再灌注期的相对F480/F420分别为缺血前的(115.27±4.52)%和(116.99±3.99)%,在再灌注期给予GSH,则活性氧含量降为缺血前的(101.14±3.20)%。结论双波长比例法可用于测定细胞内活性氧。Objective To establish the double wavelengths ratio for detecting the concentrations of reactive oxygen species（ROS） in single cell.Methods ①Adult rat cardiomyocytes were isolated by enzymatic dissociation.Fluorescence signals of the single cell were recorded by the ion imaging system for finding the two excitation wavelengths which made the biggest and the smallest fluorescence signal intensities.②Cardiomyocytes were perfused with BPS-4 to imitate the ischemia/reperfusion model.Some cardiomyocytes were reperfused with GSH.The intracellular ROS concentrations were measured.Results ①The values of the fluorescence intensity at 480 nm and 420 nm were the biggest and smallest,respectively.F480/F420 was used to represent the intracellular ROS concentrations.②At mimic ischemia and reperfusion stages,intracellular ROS fluorescence intensities（F480/F420） increased progressively to（115.27±4.52）% and（116.99±3.99）%of pre-ischemia respectively.When GSH was used in reperfusion,ROS fluorescence intensity decreased quickly to（101.14±3.20）% of pre-ischemia.Conclusion Double wavelengths ratio could be used to detect intracellular ROS.

关 键 词：活性氧 缺血再灌注 还原型谷胱甘肽 

分 类 号：O657[理学—分析化学]