纳米二氧化硅对正常人外周血单个核细胞凋亡的诱导作用  

Induction of silica nanoparticles on apoptosis of normal human peripheral blood mononuclear cells

作　　者：钟建斌[1] 雷泽林[2] 唐艳[3] 张丹[1] 刘举科[4] 马建秀[1] 

机构地区：[1]西北民族大学医学院,甘肃兰州730030 [2]兰州大学第一医院呼吸科,甘肃兰州730000 [3]兰州大学第一医院中心实验室,甘肃兰州730000 [4]兰州城市学院,甘肃兰州730070

出　　处：《中国药理学与毒理学杂志》2016年第11期1176-1181,共6页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(31260133);西北民族大学2015年国家级大学生创新创业训练计划项目(201510742088)~~

摘　　要：目的观察纳米二氧化硅(SiO_2)对正常人外周血单个核细胞(PBMC)凋亡的诱导作用,探讨纳米SiO_2对人体的毒理特性。方法透射电子显微镜观察纳米SiO_2粒径、形态及分散性;采集6名健康志愿者外周血标本,进行体外全血培养和体外PBMC分离培养;纳米SiO_212.5,25,50,100和200 mg·L^(-1)与外周全血细胞和PBMC作用24 h后,血常规法检测外周血白细胞数目,罗丹明标记鬼笔环肽标记法于荧光倒置显微镜观察PBMC骨架微丝结构,AnnexinⅤ-FITC/PI双染检测细胞早期凋亡率,ELISA检测细胞培养上清液活性氧(ROS)自由基水平。结果与正常对照组相比,纳米SiO_2100和200 mg·L^(-1)引起外周全血细胞中淋巴细胞和单核细胞数目明显减少(P<0.01);PBMC胞体皱缩、变圆,细胞核固缩并出现空泡化,细胞骨架微丝结构破坏;早期凋亡率和胞内ROS水平明显升高(P<0.01),且细胞早期凋亡率与胞内ROS水平呈明显的正相关(R^2=0.847,P<0.01)。结论纳米SiO_2对PBMC有明显的细胞毒性作用,可诱导细胞凋亡;激发氧化应激反应可能是其导致细胞凋亡的主要原因。OBJECTIVE To elucidate the toxicological properties of silica nanoparticles（nanoSiO_2）by investigating their effect on human peripheral blood mononuclear cells（PBMCs）. METHODS The particle size,morphology and dispersion of nano- SiO_2 were characterized by the transmission electron microscope. Human blood samples were provided by 6 healthy volunteers,while PBMCs were isolated from the peripheral blood cells and cultured in vitro. The cells were treated with 10 nm nano-SiO_2 in RPMI 1640 media at the concentration of 0（normal control）,12.5,25,50,100 and200 mg·L^（-1）for 24 h. Blood routine examination was performed. The human PBMC morphology was observed under an inverted microscope,followed by detection and analysis of the microfilament using TRITC-phallodin and fluorescence microscopy,respectively. After staining with AnnexinⅤ-FITC and propidumiodide,the cell early apoptosis rate was measured by flow cytometry. Reactive oxygen species（ROS）production in the supernatant medium liquid was measured with ELISA. RESULTS Compared with normal control group,the proportion of lymphocytes and macrophages significantly decreased when the cells were treated with nano- SiO_2100 and 200 mg · L^（-1）. The cell morphology changed and the microfilament was disrupted. The early apoptosis rate and intracellular ROS level both significantly increased along with the nano-SiO_2 exposure concentration（P〈0.01）. In addition,there was a significant positive correlation between ROS and apoptosis rate（R^2=0.847,P〈0.01）. CONCLUSION Nano-SiO_2 demonstrates cytotoxicity when exposed to PBMCs. The elevated level of oxidative stress is probably a major reason for early apoptosis.

关 键 词：纳米二氧化硅 外周血 单个核细胞 氧化应激 细胞凋亡 

分 类 号：R114[医药卫生—卫生毒理学]