纳米二氧化硅对BEAS-2B细胞的毒性研究

发布时间：2018-05-06 13:43

本文选题：纳米二氧化硅 + 支气管上皮细胞　；参考：《毒理学杂志》2015年03期

【摘要】：目的评价纳米二氧化硅颗粒物对人正常支气管上皮细胞(BEAS-2B)的毒性作用及可能机制。方法(1)扫描电子显微镜(Scanning Electron Microscope,SEM)观察经100μg/ml的20 nm二氧化硅和结晶型二氧化硅(MinU-Sil 5)暴露24~72 h后的BEAS-2B细胞形态变化;(2)应用3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)比色法检测实验组(纳米二氧化硅处理组)、阳性对照组(Min-U-Sil 5处理组)的细胞活力变化,分别测定不同实验条件下的还原型谷胱甘肽(GSH)、细胞内活性氧(ROS)和乳酸脱氢酶(LDH)水平;(3)流式细胞仪分析经暴露不同浓度20 nm二氧化硅后的细胞周期变化。结果 (1)经20 nm二氧化硅刺激24 h后BEAS-2B细胞体积增大,胞浆疏松,细胞数量减少,并随刺激时间延长而加重,阳性对照组上述变化不明显;(2)BEAS-2B细胞活力随暴露浓度和暴露时间的增加而下降,暴露浓度50μg/ml时20 nm二氧化硅组的细胞活力下降与阳性对照组间比较,差异有统计学意义(P0.05),暴露浓度为100和200μg/ml时20 nm组与50 nm组间比较差异有统计学意义(P0.05)。100μg/ml浓度暴露24 h后,20 nm二氧化硅组细胞活力下降情况与阳性对照组间比较,差异有统计学意义(P0.05),暴露72 h后各组间细胞活力下降差异均有统计学意义(P0.05)。BEAS-2B细胞培养基中的细胞内活性氧(ROS)升高、还原型谷胱甘肽(GSH)下降和乳酸脱氢酶(LDH)升高,并随暴露时间延长和二氧化硅粒径减小而变化;(3)纳米二氧化硅可使G2/M期停滞和G1期细胞增多,并呈现浓度效应。结论纳米二氧化硅造成BEAS-2B细胞的细胞毒性作用强于结晶型二氧化硅(Min-U-Sil 5),纳米二氧化硅对BEAS-2B细胞的细胞毒性作用,具有浓度-效应、时间-效应和粒径-效应现象,细胞毒性作用并最终引起细胞氧化损伤。
[Abstract]:Objective to evaluate the toxic effect of nano-silica particles on human normal bronchial epithelial cells (BEAS-2B) and its possible mechanism. Methods 1) scanning electron microscope (SEM) scanning electron microscopy (SEM) was used to observe the morphological changes of BEAS-2B cells after exposure to 100 渭 g/ml 20 nm silica and crystalline silica (MinU-Sil 5) for 2472 h). The 3-tir 45-dimethylthiazol-2-ylthiazol-2-yl-25-diphenyltetrazolium bromidedet assay was used to detect the morphological changes of BEAS-2B cells in the experimental group (nano-silica treated group). The change of cell activity in the positive control group (Min-U-Sil 5 treatment group), The levels of reduced glutathione (GSH), intracellular reactive oxygen species (Ros) and lactate dehydrogenase (LDH) were measured by flow cytometry (FCM) to analyze the cell cycle changes after exposure to 20 nm silica at different concentrations. Results 1) after being stimulated with 20 nm silica for 24 h, the volume of BEAS-2B cells increased, the cytoplasm became loose and the number of cells decreased, and the number of cells increased with the prolongation of stimulation time. The above changes were not obvious in the positive control group. The activity of BEAS-2B cells decreased with the increase of the exposure concentration and exposure time. The cell viability of the 20nm silica group at the exposure concentration of 50 渭 g/ml was lower than that of the positive control group. There was significant difference between the 20 nm group and the 50 nm group when the exposure concentration was 100 渭 g/ml and 200 渭 g/ml. The cell viability of the 20 nm silica group was decreased after 24 h exposure to P0.05 + 100 渭 g/ml concentration, compared with that of the positive control group. The difference was statistically significant (P 0.05). After 72 h exposure, there were significant differences in the decrease of cell viability and the increase of intracellular reactive oxygen species (Ros), reduced glutathione (GSH) and lactate dehydrogenase (LDH) in BEAS-2B cell culture medium. With the prolongation of the exposure time and the decrease of the particle size of silica, the nano silica can make the G 2 / M phase arrest and G1 phase cells increase, and present a concentration effect. Conclusion the cytotoxicity of BEAS-2B cells induced by nano-silica is stronger than that of crystallized silica Min-U-Sil 5G. The effects of nano-silica on BEAS-2B cells are concentration-effect, time-effect and particle size effect. Cytotoxicity and ultimately cell oxidative damage.
【作者单位】：昆明医科大学第三附属医院/云南省肿瘤医院胸外一科;昆明医科大学第三附属医院/云南省肿瘤医院肿瘤研究所;
【分类号】：R994.1

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