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DNA氧化损伤作为检测二氧化钛纳米颗粒诱导的遗传毒性的敏感遗传终点。

DNA Oxidative Damage as a Sensitive Genetic Endpoint to Detect the Genotoxicity Induced by Titanium Dioxide Nanoparticles.

作者信息

Chen Zhangjian, Shi Jiaqi, Zhang Yi, Han Shuo, Zhang Jiahe, Jia Guang

机构信息

Department of Occupational and Environmental Health Sciences, School of Public Health, Peking University, Beijing 100191, China.

Beijing Key Laboratory of Toxicological Research and Risk Assessment for Food Safety, Peking University, Beijing 100191, China.

出版信息

Nanomaterials (Basel). 2022 Jul 29;12(15):2616. doi: 10.3390/nano12152616.

Abstract

The genotoxicity of nanomaterials has attracted great attention in recent years. As a possible occupational carcinogen, the genotoxic effects and underlying mechanisms of titanium dioxide nanoparticles (TiO NPs) have been of particular concern. In this study, the effect of TiO NPs (0, 25, 50 and 100 µg/mL) on DNA damage and the role of oxidative stress were investigated using human bronchial epithelial cells (BEAS-2B) as an in vitro model. After detailed characterization, the cytotoxicity of TiO NPs was detected. Through transmission electron microscopy (TEM), we found that TiO NPs entered the cytoplasm but did not penetrate deep into the nucleus of cells. The intracellular levels of reactive oxygen species (ROS) significantly increased in a dose-dependent manner and the ratios of GSH/GSSG also significantly decreased. The results of the normal comet assay were negative, while the Fpg-modified comet assay that specifically detected DNA oxidative damage was positive. Meanwhile, -acetyl-L-cysteine (NAC) intervention inhibited the oxidative stress and genotoxicity induced by TiO NPs. Therefore, it was suggested that TiO NPs could induce cytotoxicity, oxidative stress and DNA oxidative damage in BEAS-2B cells. DNA oxidative damage may be a more sensitive genetic endpoint to detect the genotoxicity of TiO NPs.

摘要

近年来,纳米材料的遗传毒性引起了极大关注。作为一种可能的职业致癌物,二氧化钛纳米颗粒(TiO NPs)的遗传毒性效应及潜在机制一直备受关注。在本研究中,以人支气管上皮细胞(BEAS-2B)作为体外模型,研究了TiO NPs(0、25、50和100 µg/mL)对DNA损伤的影响以及氧化应激的作用。在详细表征后,检测了TiO NPs的细胞毒性。通过透射电子显微镜(TEM),我们发现TiO NPs进入了细胞质,但未深入细胞核。细胞内活性氧(ROS)水平以剂量依赖方式显著增加,谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)比值也显著降低。正常彗星试验结果为阴性,而专门检测DNA氧化损伤的Fpg修饰彗星试验结果为阳性。同时,N-乙酰-L-半胱氨酸(NAC)干预抑制了TiO NPs诱导的氧化应激和遗传毒性。因此,提示TiO NPs可诱导BEAS-2B细胞的细胞毒性、氧化应激和DNA氧化损伤。DNA氧化损伤可能是检测TiO NPs遗传毒性更敏感的遗传终点。

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