体内外遗传毒性实验组合评价纳米银的遗传毒性

发布时间：2018-05-14 17:25

本文选题：纳米银 + 小鼠淋巴瘤细胞实验　；参考：《军事医学》2015年12期

【摘要】：目的采用小鼠淋巴瘤细胞tk+/-位点突变实验、大鼠肝细胞碱性彗星电泳实验以及大鼠外周血网织红细胞微核实验观察纳米银的体内外遗传毒性。方法 (1)以2.5、5.0、10.0、20.0、40.0和80.0μg/ml浓度纳米银处理L5178Y细胞3 h,确定细胞毒性水平;再以0.63、1.25、2.50和5.00μg/ml浓度处理细胞3 h,分析纳米银致基因突变频率。(2)SD大鼠单次尾静脉注射纳米银0.63、1.25、2.50、5.00 mg/kg,24 h后分别采用碱性彗星电泳和流式细胞术检测肝细胞DNA损伤和外周网织红细胞血微核形成率。结果 (1)L5178Y细胞相对悬浮增殖率随纳米银浓度的增加而降低,在5.0μg/ml时已降至阴性对照组的11.42%;在测试浓度下,tk+/-基因位点突变频率呈浓度依赖性升高,与阴性对照组差异显著(P0.05)。同时,小集落比例较阴性对照组均有显著提升(P0.01)。(2)纳米银5.0 mg/kg组给药后10 min内2只大鼠死亡,至24 h时死亡数增至3只,样本数不足不能参与统计。与阴性对照组相比,1.25、2.5 mg/kg剂量组尾部DNA百分含量均有显著性差异(P0.05),0.63 mg/kg组未见明显异常。与阴性组相比,纳米银0.63、1.25、2.5 mg/kg 3个剂量组诱发外周血网织红细胞微核发生率均有显著性差异(P0.05),并与暴露剂量呈正相关(Pearson r=0.98,P0.05)。结论实验条件下检测出纳米银具有诱发诸如基因突变、染色体畸变和原发性DNA损伤等体内外遗传毒性的潜在风险。
[Abstract]:Objective to observe the in vitro and in vivo genotoxicity of silver nanoparticles by tk /-site mutation assay in mouse lymphoma cells, alkaline comet electrophoresis in rat hepatocytes and micronucleus assay of rat peripheral blood reticulocyte. Methods 1) L5178Y cells were treated with silver nanoparticles (2.5 渭 g/ml, 10.0 渭 g/ml and 80 渭 g/ml) for 3 h to determine the cytotoxicity level. After the cells were treated with 0.63 渭 g/ml and 5.00 渭 g/ml for 3 h, the DNA damage and peripheral reticulum of hepatocytes were detected by alkaline comet electrophoresis and flow cytometry respectively after single caudal vein injection of silver nano-silver in SD rats at the dose of 0.63 ~ 1.25 ~ 2.50 mg 路kg ~ (-1) 路kg ~ (-1) ~ (-1) 路kg ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ 2 ~ (-1) ~ (-1) ~ (-1), respectively. The rate of micronucleus formation in the blood of diutrocyte. Results the relative suspension proliferation rate of L5178Y cells decreased with the increase of the concentration of silver nanoparticles, and decreased to 11.42 of the negative control group at 5.0 渭 g/ml, and the mutation frequency of tk ~ (-) gene locus increased in a concentration dependent manner at the test concentration, and the difference was significant (P 0.05) compared with the negative control group. At the same time, the small colony ratio was significantly increased compared with the negative control group (P 0.01. 0. 2) the death rate of 2 rats in the 5. 0 mg/kg group was increased within 10 min after administration, and the death rate increased to 3 at 24 h. The number of samples was not enough to participate in the statistics. Compared with the negative control group, there was significant difference in the percentage content of DNA in tail of the 1.25U 2.5 mg/kg group. There was no obvious abnormality in the 0.63 mg/kg group. Compared with the negative group, the incidence of micronucleus in peripheral blood reticulocyte (PBMC) was significantly different among the three doses of silver nanoparticles (0.63C 1.252.5 mg/kg), and there was a positive correlation with the exposure dose (P 0.05). Conclusion under experimental conditions, silver nanoparticles have the potential risk of inducing genetic toxicity in vivo and in vitro, such as gene mutation, chromosome aberration and primary DNA damage.
【作者单位】：烟台大学药学院;军事医学科学院毒物药物研究所国家北京药物安全评价研究中心抗毒药物与毒理学国家重点实验室;
【基金】：国家重大新药创制科技重大专项资助项目(2013ZX09302303,2012ZX09301-001-008)
【分类号】：R114

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