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Involvement of reactive oxygen species and high-voltage-activated calcium currents in nanoparticle zinc oxide-induced cytotoxicity in vitro

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Abstract

This study was to determine the possible neurotoxicity and mechanisms underlying the effects of nano-ZnO with sizes of 20–80 nm on central nervous system (CNS). The cytotoxicity of nano-ZnO was investigated in PC12 cells. The viability of cells was observed by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the generation of reactive oxygen species (ROS) for cells was evaluated by a fluorometry assay. The apoptosis of cells was detected and analyzed by flow cytometry. In addition, effects of nano-ZnO on the properties of high-voltage-activated (HVA) calcium currents were studied in acutely isolated rat hippocampal pyramidal neurons using the whole-cell patch clamp technique. The results of MTT assay showed that nano-ZnO (10−4 g/mL) caused a significant decrease in cell viability (P < 0.05). Nano-ZnO induced intracellular accumulation of ROS and the apoptosis of PC12 cells with the increasing concentration of nano-ZnO in flow cytometric assay (P < 0.05). Further results of electrophysiological recording indicated that 10−4 g/mL nano-ZnO first altered the current–voltage curve and the peak amplitudes of HVA calcium currents at 10 min of the recording, and the peak current amplitudes were increased significantly at the end of 30 min (P < 0.05). All these results suggested that the increase of intracellular ROS was one of potential mechanisms of cellular apoptosis induced by nano-ZnO. Nano-ZnO could cause the elevation of cytosolic calcium levels by enhancement of HVA calcium currents, which would increase the generation of intracellular ROS, and consequently promote the neuronal apoptosis.

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Acknowledgments

This study was partly supported by the National Natural Science Foundation of China (31070890) and the Tianjin Research Program of Application Foundation and Advanced Technology (10JCZDJC19100).

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Authors and Affiliations

  1. College of Medicine, Nankai University, Tianjin, 300071, China

    Jingxia Zhao, Shichang Liu & Zhuo Yang

  2. Tianjin Medical University, Tianjin, 300070, China

    Jingxia Zhao

  3. Tianjin First Central Hospital, Tianjin, 300192, China

    Yang Yao

  4. College of Life Science, Nankai University, Tianjin, 300071, China

    Tao Zhang

  5. Science and Technology Research Institute, University of Hertfordshire, Hatfield, AL10 9AB, UK

    Guogang Ren

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  1. Jingxia Zhao
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  2. Yang Yao
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  3. Shichang Liu
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  4. Tao Zhang
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  6. Zhuo Yang
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Correspondence to Zhuo Yang.

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Zhao Jingxia and Yao Yang contributed equally to this study.

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Zhao, J., Yao, Y., Liu, S. et al. Involvement of reactive oxygen species and high-voltage-activated calcium currents in nanoparticle zinc oxide-induced cytotoxicity in vitro. J Nanopart Res 14, 1238 (2012). https://doi.org/10.1007/s11051-012-1238-1

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