J4 ›› 2011, Vol. 37 ›› Issue (2): 236-240.

• 基础研究 • 上一篇    下一篇

纳米SiO2颗粒在HepG2细胞中的分布及其细胞毒性

孙磊1|李阳1|刘晓梅1|杜忠君1|金明华1|孙志伟1|2   

  1. 1. 吉林大学公共卫生学院卫生毒理学教研室|吉林 长春 130021;2. 首都医科大学公共卫生与家庭医学学院卫生毒理与卫生化学系| 北京 100069
  • 收稿日期:2010-12-08 出版日期:2011-03-28 发布日期:2011-03-28
  • 通讯作者: 孙志伟(Tel:010-83911507,E-mail:zwsun@ccmu.edu.cn) E-mail:zwsun@ccmu.edu.cn
  • 作者简介:孙 磊(1977-)|男|吉林省长春市人|在读医学博士|主要从事纳米材料的卫生毒理学方面的究。

Distribution |of silica nanoparticles in HepG2cells and its relationship with |cytotoxicity

SUN Lei1| LI Yang1|LIU Xiao-mei1| DU Zhong-jun1| JIN Ming-hua1| SUN Zhi-wei1,2
  

  1. 1. Department of Health Toxicology| School of Pubilc Health| Jilin University| Changchun 130021|China;2. Department of Toxicology and Sanitary Chemistry,School of Public Health and Family Medicine,Capital Medical University,Beijing 100069,China
  • Received:2010-12-08 Online:2011-03-28 Published:2011-03-28
  • Supported by:

     教育部高等学校博士学科点专项科研基金资助课题 (20090061110062);科学前沿与交叉学科创新项目资助课题 (200903112)

摘要:

目的:检测纳米SiO2颗粒作用于HepG2细胞后纳米颗粒的细胞摄取、分布情况及颗粒对细胞的毒性作用,初步探讨纳米颗粒的摄取、分布与细胞毒性的关系。方法:采用透射电镜(TEM)及动态光散射法检测纳米SiO2颗粒的表征;纳米SiO2颗粒作用HepG2细胞后,用荧光显微镜及TEM观察纳米SiO2颗粒的细胞摄取及细胞内分布情况;不同浓度纳米SiO2颗粒(0、25、50、100 及200 mg·-1)作用于HepG2细胞24 h后,采用MTT法检测细胞存活率,用Rhodamine123标记流式细胞术检测线粒体膜电位变化。结果:TEM结果显示,纳米SiO2颗粒呈球形,分散性好,大小均一。动态光散射法结果显示,纳米SiO2颗粒在无血清DMEM中粒径约为94 nm,分散
性较好。纳米SiO2颗粒作用于HepG2细胞3 h即可进入细胞;作用24 h后,可以单一颗粒或成簇的形态分散在胞质内,并在线粒体等细胞器内沉积;不同浓度纳米SiO2颗粒作用于细胞24 h后,各组细胞存活率及线粒体膜电位较对照组均有显著下降(P<0.05),且随着剂量增加细胞存活率和线粒体膜电位降低。结论:纳米SiO2颗粒进入细胞并在细胞器中沉积,可导致线粒体等细胞器的损伤,进而产生细胞毒性作用,抑制细胞增殖。

关键词: 纳米SiO2颗粒;细胞内分布;线粒体;细胞毒性

Abstract:

Abstract:Objective To detect the cell uptake and intracellular distribution of silica nanoparticles in HepG2 cells after treated with silica nanoparticles and the cytotoxicity of HepG2 cells caused by the particles and discuss the relationship between intracellular distribution of nanoparticles and cytotoxicity.
Methods The size,shape and dispersibility of silica nanoparticles were detected by transmission electron microscope (TEM) and dynamic light scattering method. The cell uptake and intracellular distribution of silica nanoparticles were observed by fluorescence microscope and TEM. The HepG2 cells were exposed to different concentrations (0,25,50,100 and 200 mg·L-1) of silica nanoparticles for 24 h, the cell viability was reflected by MTT assay and the changes of mitochondrial membrane potential (MMP) were measured by flow cytometry. Results The TEM results  showed that silica nanoparticles were spherical and well dispersed. The dynamic light scattering method results manifested that the size of silica nanoparticles in serum-free DMEM was about 94 nm and well dispersed. Silica nanoparticles could enter into the cells after HepG2 cells were treated with the particles for 3 h. After 24 h treatment,silica nanoparticles were found to disperse in cytoplasm with single particle or cluster form. The particles coulddeposit in some organelles such as mitochondria. After the HepG2 cells were exposed to different concentrations of silica nanoparticles for 24 h,the cell viabiliyand MMP in treated groups were decreased significantly (P<0.05) in a dose-dependent manner. Conclusion The silica nanoparticles can damage the organelles which is one of the mechanisms of cytotoxicity induced by silica nanoparticles.

Key words: silica nanoparticles;intracellular distribution;mitochondria;cytotoxicity

中图分类号: 

  • R994