纳米二氧化硅颗粒对肝HL-7702细胞的毒性作用

doi:10.13481/j.1671-587x.20150501

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (05): 885-890.doi: 10.13481/j.1671-587x.20150501

• 基础研究 • 下一篇

纳米二氧化硅颗粒对肝HL-7702细胞的毒性作用

潘涛^1,2, 郭彩霞³, 金明华¹, 刘晓梅¹, 刘颖¹, 杜海英¹, 孙志伟^1,4

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. 西安医学院公共卫生系, 陕西西安 710021;
3. 首都医科大学公共卫生学院劳动卫生与环境卫生学系, 北京 100069;
4. 首都医科大学公共卫生学院卫生化学与毒理学系, 北京 100069

收稿日期:2014-10-06 出版日期:2015-09-28 发布日期:2015-09-29
通讯作者: 孙志伟,教授,博士研究生导师(Tel:010-83911507,E-mail:zwsun@hotmail.com) E-mail:zwsun@hotmail.com
作者简介:潘涛(1984-),男,山东省日照市人,医学硕士,主要从事环境毒理学方面的研究。
基金资助:
国家自然科学基金资助课题(81172704)

Cytotoxicity of SiO₂ nanoparticles on liver HL-7702 cells

PAN Tao^1,2, GUO Caixia³, JIN Minghua¹, LIU Xiaomei¹, LIU Ying¹, DU Haiying¹, SUN Zhiwei^1,4

1. Department of Health Toxicology, School of Public Health, Jilin University, Changchun 130021, China;
2. Department of Public Health, Xi'an Medical University, Xi'an 710021, China;
3. Department of Occupational and Environmental Health, School of Public Health, Capital Medical University, Beijing 100069, China;
4. Department of Sanitary Chemistry and Toxicology, School of Public Health, Capital Medical University, Beijing 100069, China

Received:2014-10-06 Online:2015-09-28 Published:2015-09-29

摘要/Abstract

摘要：

目的:探讨纳米二氧化硅(SiO₂)颗粒对肝细胞HL-7702的毒性作用,阐明其作用机制。方法:体外培养的人正常HL-7702肝细胞随机分为阴性对照组和12.5、25.0、50.0、100.0 mg·L^-1 SiO₂组。采用透射电子显微镜(TEM)和动态光散射(DLS)法对纳米SiO₂颗粒进行表征和粒径检测。细胞处理 24 h 后,HE染色观察各组细胞形态学;MTT实验检测各组细胞存活率;乳酸脱氢酶(LDH)活力实验检测各组细胞培养液中LDH活力;流式细胞术(FCM)检测各组细胞中活性氧(ROS)水平。结果:TEM观察,纳米SiO₂呈球形,颗粒大小较均匀一致,分散性较好,颗粒平均粒径为(65.87±9.02)nm;DLS法检测,纳米SiO₂颗粒在高纯水和RPMI-1640培养液中的水合粒径明显增大,分别为(124.57±8.02)和(139.32±9.93)nm。HE染色,纳米SiO₂颗粒能够导致HL-7702细胞形态改变,25.0和50.0 mg·L^-1 SiO₂组细胞数目减少,排列紊乱;100.0 mg·L^-1 SiO₂组细胞质皱缩、细胞排列稀疏,部分细胞呈凋亡的形态学表现。与阴性对照组比较,各浓度纳米SiO₂组HL-7702细胞存活率均下降,50.0和100.00 mg·L^-1 SiO₂组细胞存活率明显下降(P<0.05);50.0和100.00 mg·L^-1 SiO₂组培养液中LDH 活力明显升高(P<0.05),细胞中ROS水平明显升高(P<0.05)。结论:纳米SiO₂颗粒能够对HL-7702细胞产生毒性作用,并呈一定的浓度依赖效应;其毒性作用机制可能与细胞中ROS的产生有关联。

关键词: 纳米SiO₂颗粒, HL-7702细胞, 细胞毒性, 活性氧

Abstract:

Objective To study the cytotoxicity of SiO₂ nanoparticles (NPs) on the liver HL-7702 cells,and to clarify its mechanism. Methods The in vitro cultured human normal liver HL-7702 cells were randomly divided into control group and 12.5,25.0,50.0,100.0 mg·L^-1 SiO₂ groups,respectively.Transmission electron microscope (TEM) was used to characterize the SiO₂ NPs,and dynamic light scattering(DLS) method was used to analyze the particle size of SiO₂ NPs.After 24 h exposure,the morphology of HL-7702 cells were observed by HE staining;MTT assay was used to determine the viability of HL-7702 cells;lactate dehydrogenase (LDH) release assay was used to detect the LDH activity in culture medium of HL-7702 cells;flow cytometry (FCM) was applied to examine the level of intracellular reactive oxygen species (ROS). Results The TEM results showed that the SiO₂ NPs were spherical,universally sized and more or less dispersed well,and the average particle size of SiO₂ NPs were (65.87±9.02) nm.The DLS method results manifested that the SiO₂ particle sizes of the SiO₂ NPs were (124.57±8.02) nm in high purity water and (139.32±9.93) nm in RPMI-1640 solution.The HE staining results revealed that the SiO₂ NPs could cause the morphological changes of HL-7702 cells,and the number of HL-7702 cells was reduced and the arrangement was disorder in 25.0 and 50.0 mg·L^-1 SiO₂ groups;in 100.0 mg·L^-1 SiO₂ group,the cells were shrunken and sparse,and part of the cells appeared the morphological features of apoptosis.Compared with control group,the cell viabilities in different concentrations of SiO₂ groups were decreased,especialy in 50.0 and 100.0 mg·L^-1 SiO₂ groups (P<0.05),while the LDH activities in culture medium and the intracellular ROS levels in 50.0 and 100.0 mg·L^-1 SiO₂ groups were increased (P<0.05). Conclusion SiO₂ NPs could cause the cytotoxicity on the HL-7702 cells in a concentration-dependent manner,and the cytotoxicity may be associated with the generation of intracellular ROS.

Key words: SiO₂ nanoparticles, HL-7702 cells, cytotoxicity, reactive oxygen species

中图分类号:

潘涛, 郭彩霞, 金明华, 刘晓梅, 刘颖, 杜海英, 孙志伟. 纳米二氧化硅颗粒对肝HL-7702细胞的毒性作用[J]. 吉林大学学报(医学版), 2015, 41(05): 885-890.

PAN Tao, GUO Caixia, JIN Minghua, LIU Xiaomei, LIU Ying, DU Haiying, SUN Zhiwei. Cytotoxicity of SiO₂ nanoparticles on liver HL-7702 cells[J]. Journal of Jilin University Medicine Edition, 2015, 41(05): 885-890.

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纳米二氧化硅颗粒对肝HL-7702细胞的毒性作用

Cytotoxicity of SiO₂ nanoparticles on liver HL-7702 cells

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纳米二氧化硅颗粒对肝HL-7702细胞的毒性作用

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Cytotoxicity of SiO₂ nanoparticles on liver HL-7702 cells