N-乙酰半胱氨酸对纳米二氧化硅所致细胞毒性的抑制作用

doi:10.13481/j.1671-587x.20150310

摘要/Abstract

摘要：

目的:探讨N-乙酰半胱氨酸(NAC)对纳米二氧化硅(SiO₂)颗粒导致细胞中活性氧(ROS)水平升高的抑制作用,阐明纳米SiO₂颗粒致细胞毒性的机制。方法:选用粒径为68 nm的纳米SiO₂颗粒,以体外培养的人正常肝细胞(L-02)为模型,分为对照组和不同浓度(1、2、5、10 mmol·L^-1)NAC预处理组、不同浓度(10、20、50、100 mg·L^-1)纳米SiO₂颗粒暴露组和NAC预处理的纳米SiO₂颗粒暴露组。采用MTT法测定不同浓度NAC预处理后的细胞存活率以及不同浓度NAC预处理后暴露于纳米SiO₂颗粒的细胞存活率;倒置显微镜观察NAC预处理后暴露于纳米SiO₂颗粒的细胞形态;荧光酶标仪检测NAC预处理后暴露于不同浓度纳米SiO₂颗粒的细胞中ROS水平。结果:NAC预处理组细胞存活率均高于对照组,在5 mmol·L^-1 浓度时达到峰值,但与对照组比较差异无统计学意义(P>0.05);不同浓度NAC能使暴露于纳米SiO₂颗粒的细胞存活率升高,以5 mmol·L^-1 NAC的作用最明显(P<0.05);与纳米SiO₂颗粒暴露组比较,NAC预处理的纳米SiO₂颗粒暴露组细胞数目增多,细胞边界较明显;随纳米SiO₂颗粒浓度的升高,细胞中 ROS水平随之升高,呈现剂量依赖效应,在50和100 mg·L^-1浓度时与对照组比较差异有统计学意义(P<0.05);与相同浓度纳米SiO₂颗粒暴露组比较,NAC预处理的纳米SiO₂颗粒暴露组细胞中ROS水平降低,差异有统计学意义(P<0.05)。结论:NAC在一定程度上能够抑制纳米SiO₂颗粒所致的细胞存活率下降和细胞中ROS水平升高,细胞中ROS水平升高是纳米SiO₂颗粒导致人正常肝细胞毒性的作用机制之一。

关键词: 纳米二氧化硅, 细胞毒性, N-乙酰半胱氨酸, 活性氧, 肝细胞L-02

Abstract:

Objective To detect the inhibitory effects of N-acetyl-cysteine(NAC)on the increasing of reactive oxygen species(ROS)level induced by SiO₂ nanoparticles, and to clarify the mechanism of cytotoxicity induced by SiO₂ nanoparticles. Methods The 68 nm SiO₂ nanoparticles were selected and the in vitro cultured human normal liver L-02 cells were used as cell model.The L-02 cells were divided into control group, different concentrations of NAC(1, 2, 5, 10 mmol·L^-1)pretreatment groups, different concentrations of SiO₂ nanoparticle(10, 20, 50, 100 mg·L^-1)groups and SiO₂ nanoparticle exposure with NAC pretreatment groups.MTT assay was applied to measure the viabilities of the L-02 cells in NAC pretreatment groups with and without SiO₂ nanoparticle exposure.The morphology of L-02 cells pretreated with NAC followed by SiO₂ nanoparticles exposure was observed by inverted microscope.The intracellular ROS levels were measured by fluorescence microplate reader to evaluate NAC effects towards different concentrations of SiO₂ nanoparticles. Results The cell viabilities in NAC pretreatment groups were elevated, and peaked in 5 mmol·L^-1 group.However, compared with control group, the NAC did not increase the cell viability significantly(P>0.05).Compared with SiO₂ nanoparticle exposure group without NAC, the cell viabilities in NAC pretreatment groups were increased(P<0.05).The image of NAC pretreatment group showed higher cell number and more distinct cell boundary compared with SiO₂ nanoparticle group without NAC.With the increase of SiO₂ nanoparticles concentration, the intracellular ROS levels were increased in a dose-dependent manner.Compared with ocntrol group, the ROS levels in 50 and 100 mg·L^-1 SiO₂ nanoparticle groups were increased significantly(P<0.05).The ROS levels in SiO₂ nanoparticles exposure with NAC pretreatment groups were decreased compared with SiO₂ nanoparticles exposure groups at the same concentration(P<0.05). Conclusion NAC can inhibit the decrease of cell viability and the increase of ROS level induced by SiO₂ nanoparticles to some extent.The intracellular ROS level increase is one of the mechanisms of SiO₂ nanoparticles-induced cytotoxicity.

Key words: SiO₂ nanoparticles, cytotoxicity, N-acetyl-cysteine, reactive oxygen species, liver L-02 cells

中图分类号:

耿维佳, 李阳, 于永波, 于洋, 段军超, 杨玉梅, 邹洋, 孙志伟. N-乙酰半胱氨酸对纳米二氧化硅所致细胞毒性的抑制作用[J]. 吉林大学学报(医学版), 2015, 41(03): 486-490.

GENG Weijia, LI Yang, YU Yongbo, YU Yang, DUAN Junchao, YANG Yumei, ZOU Yang, SUN Zhiwei. Inhibitory effects of N-acetyl-cysteine on SiO₂ nanoparticles-induced cytotoxicity[J]. Journal of Jilin University Medicine Edition, 2015, 41(03): 486-490.

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