张剑,
钱云霞,
宁波大学海洋学院, 宁波 315823
作者简介: 朱跃骅(1993-),男,硕士研究生,研究方向为海洋环境与水产病害研究,E-mail:15757826826@163.com.
通讯作者: 钱云霞,qianyunxia@nbu.edu.cn
基金项目: 国家重点研发计划重点专项（2016YFC1402405）中图分类号: X171.5
Toxic Effect and Mechanism of Nano-ZnO in Hepatocytes of Sciaenops ocellatus
Zhu Yuehua,Zhang Jian,
Qian Yunxia,
College of Marine Sciences, Ningbo University, Ningbo 315823, China
Corresponding author: Qian Yunxia,qianyunxia@nbu.edu.cn
CLC number: X171.5
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摘要:纳米氧化锌（ZnO NPs）的广泛应用所引发的环境与健康风险备受关注。为探究ZnO NPs对水生生物的毒性影响，以美国红鱼的原代肝细胞为实验对象，通过MTT法和中性红摄取（NRU）法检测细胞存活率，以评估ZnO NPs的细胞毒性；美国红鱼肝细胞经ZnO NPs处理6 h后，检测肝细胞内丙二醛（MDA）含量，并用流式细胞仪检测细胞对ZnO NPs的吞噬、胞内活性氧（ROS）的释放以及细胞凋亡率的变化。结果表明，美国红鱼肝细胞暴露于37.5 μg·mL-1和50 μg·mL-1 ZnO NPs 6 h后，细胞存活率较对照组显著降低，且ZnO NPs的毒性具有浓度和时间依赖性。研究表明，ZnO NPs进入细胞后，细胞内ROS产量增加，引起氧化应激，诱导细胞凋亡。
关键词: 纳米氧化锌/
美国红鱼/
细胞毒性/
氧化应激/
流式细胞
Abstract:The toxicity of zinc oxide nano particles (ZnO NPs) has received increasing attention in recent years. This experiment was aimed to assess the toxicity effects of ZnO NPs on primary cultured hepatocytes of Sciaenops ocellatus. The in vitro cytotoxicity of ZnO NPs was tested using MTT-fomazan assay and neutral red uptake (NRU) assay. Cellular uptake of ZnO NPs, intracellular reactive oxygen production and apoptosis rate were measured by flow cytometry after exposure to ZnO NPs for 6 h, and the content of malondialdehyde (MDA) were also detected by colorimetry. The results showed that the cell viability decreased after Sciaenops ocellatus hepatocytes were exposed to 37.5 μg·mL-1 and 50 μg·mL-1 ZnO NPs for 6 h. The studies indicated that ZnO NPs inhibited the growth of the hepatocytes of Sciaenops ocellatus in a concentration- and time-dependent manner. It is demonstrated that ZnO NPs induced cells apoptosis, which was mediated by oxidative stress as evidenced by an increase in intracellular ROS and the content of MDA generation.
Key words:nano zinc oxide/
Sciaenops ocellatus/
cytotoxicity/
oxidative stress/
flow cytometry.
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