摘要
目的研究3种不同磁性纳米颗粒对体外培养的血管内皮细胞中活性氧(reactive oxygen species,ROS)水平和细胞间连接的影响,探讨二者之间的关联性。方法将原代人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)随机分为对照组和不同磁性纳米颗粒暴露组。利用动态光散射(dynamic light scattering,DLS)对纳米颗粒的粒径和电势进行表征;采用细胞计数试剂盒(cell counting Kit-8,CCK-8)法测定细胞活性;通过二氯二氢荧光素-乙酰乙酸酯(2',7'-dichlorofluorescin diacetate,DCFH-DA)荧光探针标记和流式细胞术检测细胞中ROS水平;利用普鲁士蓝染色和透射电镜方法观察内皮细胞对磁性纳米颗粒的摄取。对细胞表面血管内皮钙黏蛋白(vascular endothelial cadherin,VE-cadherin)进行免疫荧光标记,在激光共聚焦显微镜下观察细胞间连接,并通过Western blot检测VE-cadherin表达水平。结果磁性纳米颗粒能诱导内皮细胞内ROS水平上升,降低VE-cadherin表达水平,细胞间缝隙增大。抗氧化剂N-乙酰半胱氨酸处理可使ROS水平下降并减少细胞缝隙。由于组分、表面修饰、尺寸等因素不同,磁性纳米颗粒对内皮细胞活性、ROS水平及VE-cadherin产生不同程度的影响。结论不同磁性纳米颗粒对内皮细胞活性氧和细胞间连接的影响不同;在实验所采用的低剂量暴露下可影响内皮细胞连接的完整性。
Objective To study the effects of three kinds of magnetic nanomaterials on reactive oxygen species,cell junction of vascular endothelial cells in vitro and their relationship. Methods Primary human umbilical vein endothelial cells（HUVECs） were randomly divided into control and magnetic nanoparticles exposed groups.Dynamic light scattering（DLS） was used to characterize the particle size and charge of nanomaterials. CCK-8 method was used to measure cell viability. Flow cytometrywas used to observe the level of ROS in cells by DCFH-DA fluorescent probe labeling. Using iron staining and transmission electron microscopy（TEM） to observe the uptake of magnetic nanomaterials by endothelial cells.The cells were fluorescently labeled with VE-cadherin antibody,and the cell junctions were observed under a laser confocal microscope. The expression of VE-cadherin was analyzed by western blot. Results Magnetic nanomaterials could increase ROS in endothelial cells,decrease VE-cadherin levels and the gap between cells increased. When treated with the antioxidant N-acetylcysteine,the level of reactive oxygen decreased,while the cell junction tends to be intact. The effects of magnetic materials on the activity of endothelial cells,ROS level,VE-cadherin expression were different due to components,surface modification,and size. Conclusions Different magnetic nanoparticles have different effects on the ROS and cell junction of endothelial cells; the exposure dose used in experiments can affect the integrity of endothelial cell junction.
作者
杨爱云
温涛
武昊安
苗玉清
孟洁
刘健
张宇
樊海明
许海燕
YANG Aiyun;WEN Tao;WU Haoan;MIAO Yuqing;MENG Jie;LIU Jian;ZHANG Yu;FAN Haiming;XU Haiyan(Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Beijing, 100005;State Key Laboratory of Bioelectronics, Jiangsu Key Laboratory for Biomaterials and Devices, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096;College of Chemistry ＆ Materials Science, Northwest University, Xi＇an 710069)
出处
《北京生物医学工程》
2018年第3期235-240,共6页
Beijing Biomedical Engineering
基金
协和青年基金和中央高校基本科研业务费专项(3332016046)
中国医学科学院与健康科技创新工程项目(CIFMS 2016-I2M-3-004)资助
关键词
磁性纳米材料
内皮细胞
活性氧
细胞连接
magnetic nanomaterial
endothelial cells
reactive oxygen species
cell junction
