纳米二氧化铈对神经细胞PC12及SH-SY5Y活力的影响

doi:10.12047/j.cjap.5810.2020.014

摘要/Abstract

摘要： 目的:探究纳米二氧化铈(CeO₂)对神经细胞PC12与SH-SY5Y活力的影响。方法:合成纳米CeO₂材料,并对其结构进行表征,性能进行评估。用不同终浓度(1、2.5、5、10、25、50、75、100、150 μg/ml)的纳米CeO₂分别处理PC12细胞与SH-SY5Y细胞24 h,使用MTT法检测其细胞活力。然后使用活性氧清除剂NAC与纳米CeO₂共孵育处理PC12细胞与SH-SY5Y细胞,并用DCFH-DA探针染色,在荧光倒置显微镜下观察各组细胞的数量及其荧光强度。对实验数据采用单因素方差(one-way ANOVA)分析。结果:纳米CeO₂处理后,PC12细胞(P＜0.01)与SH-SY5Y细胞(P＜0.01)的活力都明显下降,与对照组差异明显。DCFH-DA探针染色后,发现纳米CeO₂的浓度越高,荧光强度越强,表明有活性氧(ROS)的产生。活性氧清除剂NAC与纳米CeO₂(100 μg/ml)共同处理PC12后,荧光强度明显减弱。与25 μg/ml (P＜0.01)、50 μg/ml(P＜0.01)、75 μg/ml(P＜0.01)、100 μg/ml(P＜0.01)纳米CeO₂处理组相比,共同处理组细胞活力明显增加。结论:纳米CeO₂对神经细胞PC12与SH-SY5Y的活力有明显的抑制作用,其机制可能与ROS有关。

关键词: 纳米CeO₂, 活性氧, 神经细胞, 细胞活力, 阿尔茨海默病

Abstract: Objective: To investigate the effects of cerium oxide (CeO₂) nanoparticles on the viabilities of nerve cells PC12 and SH-SY5Y. Methods: CeO₂ nanoparticles were synthesized, structures were characterized and properties were evaluated. PC12 cells and SH-SY5Y cells were treated with CeO₂ nanoparticles at different concentrations (1, 2.5, 5, 10, 25, 50, 75, 100, 150 μg/ml) for 24 h and the cell viability was measured by MTT assay. Then PC12 cells and SH-SY5Y cells were co-treated with CeO₂and active oxygen scavenger NAC and the cells were stained with DCFH-DA probe for ROS. The number of cells and the fluorescence intensity were observed under a fluorescent inverted microscope. Differences were assessed by one-way ANOVA.Results: After treatment with CeO₂ nanoparticles, the viabilities of both PC12 cells (P＜0.01) and SH-SY5Y cells (P＜0.01) were decreased comparing with the control group. After staining with DCFH-DA probe, the fluorescence intensity of the nerve cells was enhanced depending on the concentration of CeO₂ nanoparticles suggesting that CeO₂induced the generation of reactive oxygen species (ROS). The fluorescence intensity of PC12 cells was decreased after CeO₂ nanoparticles (100 μg/ml) co-treatment with active oxygen scavenger NAC. Compared with CeO₂ nanoparticles alone at 25 μg/ml (P＜0.01), 50 μg/ml (P＜0.01), 75 μg/ml (P＜0.01), 100 μg/ml (P＜0.01), the cell viability was significantly increased after co-treatment with NAC.Conclusion: CeO₂ nanoparticles has a negative effect on the viabilities of nerve cells PC12 and SH-SY5Y, and the effect might be depend on ROS.

Key words: CeO₂ nanoparticles, reactive oxygen species, neural cells, cell viability, Alzheimer’s disease

许逸岚, 赵镘娜, 朱诗涵, 韩瑜颖, 章佳瑜, 沈铄恒, 俞樟森, 张衡. 纳米二氧化铈对神经细胞PC12及SH-SY5Y活力的影响[J]. 中国应用生理学杂志, 2020, 36(1): 62-66.

XU Yi-lan, ZHAO Man-na, ZHU Shi-han, HAN Yu-ying, ZHANG Jia-yu, SHEN Shuo-heng, YU Zhang-sen, ZHANG Heng. Effects of CeO₂ nanoparticles on the viabilities of neural PC12 and SH-SY5Y cells[J]. CJAP, 2020, 36(1): 62-66.

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