Effect of ZnO nanoparticle on cell viability, zinc uptake efficiency, and zinc transporters gene expression: a comparison with ZnO and ZnSO4
X. Zhang, Z. Wang, L. Mao, X. Dong, Q. Peng, J. Chen, C. Tan, R. Huhttps://doi.org/10.17221/15/2016-CJASCitation:Zhang X., Wang Z., Mao L., Dong X., Peng Q., Chen J., Tan C., Hu R. (2017): Effect of ZnO nanoparticle on cell viability, zinc uptake efficiency, and zinc transporters gene expression: a comparison with ZnO and ZnSO4. Czech J. Anim. Sci., 62: 32-41.
Zinc plays an important role in functional and structural integrity of cells. The aim of the current study was to compare cell viability, zinc uptake efficiency, and gene expression of metallothionein (MT), divalent metal transporter (DMT-1), and other important zinc transporters (ZnTs) under experimental treatment of TPEN (N, N, N', N'-Tetrakis (2-pyridylmethyl) ethylenediamine) (2 µM), and three zinc sources (zinc oxide nanoparticle (nano-ZnO), bulk zinc oxide (ZnO), and zinc sulfate (ZnSO4)) at different levels (25, 50, and 100 µM) in rat intestinal epithelial cell line IEC-6. Cells were classified into TPEN group and TPEN + zinc sources groups. In the present study, significantly decreased cell viability was observed in TPEN group, while supplementations with nano-ZnO at all levels and ZnO (50 and 100 µM) significantly increased the cell viability. ZnSO4 at a high concentration (100 µM) inhibited cell viability. Furthermore, cells of nano-ZnO group showed the highest viability at a 25 µM concentration. The uptake efficiency of nano-ZnO is higher than that of ZnSO4 and ZnO. Additionally, a significant down-regulation for ZnT-1, ZnT-4, MT, DMT-1 mRNA with TPEN treatment was detected. Compared with the unchanged ZnT-4, all zinc treatments up-regulated the gene expressions of ZnT-1, ZnT-5, ZnT-7, MT, and DMT-1. Our results indicate that nano-ZnO is more effective than ZnO and ZnSO4 in enhancing cell viability, and its lower cytotoxicity, higher uptake efficiency, and comparative transportation at low concentration also favour its potential use as a new zinc source in feed additives.Keywords:
nano-ZnO, bulk ZnO, zinc transportation, TPEN, IEC-6 cellsReferences:
Cao J., Bobo J.A., Liuzzi J.P., Cousins R.J. (2001): Effects of intracellular zinc depletion on metallothionein and ZIP2 transporter expression and apoptosis. Journal of Leukocyte Biology, 70, 559–566.Case C. L., Carlson M. S. (2002): Effect of feeding organic and inorganic sources of additional zinc on growth performance and zinc balance in nursery pigs. Journal of Animal Science, 80, 1917- https://doi.org/10.2527/2002.8071917xCherian M (): Contemporary Issues in Toxicology Role of Metallothionein in Carcinogenesis. Toxicology and Applied Pharmacology, 126, 1-5 https://doi.org/10.1006/taap.1994.1083Cousins R.J., McMahon R.J. (2000): Integrative aspects of zinc transporters. The Journal of Nutrition, 130, 1384–1387.Cousins R. J., Liuzzi J. P., Lichten L. A. (): Mammalian Zinc Transport, Trafficking, and Signals. Journal of Biological Chemistry, 281, 24085-24089 https://doi.org/10.1074/jbc.R600011200Cragg R A (2005): Homeostatic regulation of zinc transporters in the human small intestine by dietary zinc supplementation. Gut, 54, 469-478 https://doi.org/10.1136/gut.2004.041962Davis S.R., McMahon R.J., Cousins R.J. (1998): Metallothionein knockout and transgenic mice exhibit altered intestinal processing of zinc with uniform zinc-dependent zinc transporter-1 expression. The Journal of Nutrition, 128, 825–831.De Berardis Barbara, Civitelli Gabriele, Condello Maria, Lista Pasquale, Pozzi Roberta, Arancia Giuseppe, Meschini Stefania (2010): Exposure to ZnO nanoparticles induces oxidative stress and cytotoxicity in human colon carcinoma cells. Toxicology and Applied Pharmacology, 246, 116-127 https://doi.org/10.1016/j.taap.2010.04.012Gurusamy K. S., Farooqui N., Loizidou M., Dijk S., Taanman J. W., Whiting S., Farquharson M. J., Fuller B. J., Davidson B. R. (2011): Influence of zinc and zinc chelator on HT-29 colorectal cell line. BioMetals, 24, 143-151 https://doi.org/10.1007/s10534-010-9382-5Hanley Cory, Layne Janet, Punnoose Alex, Reddy K M, Coombs Isaac, Coombs Andrew, Feris Kevin, Wingett Denise (2008): Preferential killing of cancer cells and activated human T cells using ZnO nanoparticles. Nanotechnology, 19, 295103- https://doi.org/10.1088/0957-4484/19/29/295103Hoadley J.E., Leinart A.S., Cousins R.J. (1987): Kinetic analysis of zinc uptake and serosal transfer by vascularly perfused rat intestine. American Journal of Physiology – Gastrointestinal and Liver Physiology, 252, G825–G831.Hojberg O., Canibe N., Poulsen H. D., Hedemann M. S., Jensen B. B. (): Influence of Dietary Zinc Oxide and Copper Sulfate on the Gastrointestinal Ecosystem in Newly Weaned Piglets. Applied and Environmental Microbiology, 71, 2267-2277 https://doi.org/10.1128/AEM.71.5.2267-2277.2005Huang Liping, Tepaamorndech Surapun (2013): The SLC30 family of zinc transporters – A review of current understanding of their biological and pathophysiological roles. Molecular Aspects of Medicine, 34, 548-560 https://doi.org/10.1016/j.mam.2012.05.008Kambe T. (): Cloning and Characterization of a Novel Mammalian Zinc Transporter, Zinc Transporter 5, Abundantly Expressed in Pancreatic beta Cells. Journal of Biological Chemistry, 277, 19049-19055 https://doi.org/10.1074/jbc.M200910200Kirschke C. P. (): ZnT7, a Novel Mammalian Zinc Transporter, Accumulates Zinc in the Golgi Apparatus. Journal of Biological Chemistry, 278, 4096-4102 https://doi.org/10.1074/jbc.M207644200Liuzzi J.P., Blanchard R.K., Cousins R.J. (2001): Differential regulation of zinc transporter 1, 2, and 4 mRNA expression by dietary zinc in rats. The Journal of Nutrition, 131, 46–52.Mavromichalis I, Peter C M, Parr T M, Ganessunker D, Baker D H (2000): Growth-promoting efficacy in young pigs of two sources of zinc oxide having either a high or a low bioavailability of zinc.. Journal of Animal Science, 78, 2896- https://doi.org/10.2527/2000.78112896xMiller H.M., Toplis P., Slade R.D. (2009): Can outdoor rearing and increased weaning age compensate for the removal of in-feed antibiotic growth promoters and zinc oxide? Livestock Science, 125, 121–131.Moreau J. W., Weber P. K., Martin M. C., Gilbert B., Hutcheon I. D., Banfield J. F. (2007): Extracellular Proteins Limit the Dispersal of Biogenic Nanoparticles. Science, 316, 1600-1603 https://doi.org/10.1126/science.1141064Owusu-Asiedu A., Nyachoti C. M., Marquardt R. R. (2003): Response of early-weaned pigs to an enterotoxigenic (K88) challenge when fed diets containing spray-dried porcine plasma or pea protein isolate plus egg yolk antibody, zinc oxide, fumaric acid, or antibiotic. Journal of Animal Science, 81, 1790- https://doi.org/10.2527/2003.8171790xPalmiter R. D. (): Protection against zinc toxicity by metallothionein and zinc transporter 1. Proceedings of the National Academy of Sciences, 101, 4918-4923 https://doi.org/10.1073/pnas.0401022101Palmiter Richard D., Huang Liping (2004): Efflux and compartmentalization of zinc by members of the SLC30 family of solute carriers. Pfl�gers Archiv European Journal of Physiology, 447, 744-751 https://doi.org/10.1007/s00424-003-1070-7Qin Yan, Thomas Dustin, Fontaine Charles P., Colvin Robert A. (2009): Silencing of ZnT1 reduces Zn2+ efflux in cultured cortical neurons. Neuroscience Letters, 450, 206-210 https://doi.org/10.1016/j.neulet.2008.11.069Sakabe Isamu, Paul Sharan, Dansithong Warunee, Shinozawa Takao (1998): Induction of Apoptosis in Neuro-2A Cells by Zn2+ Chelating.. Cell Structure and Function, 23, 95-99 https://doi.org/10.1247/csf.23.95Sandoval M, Henry P R, Ammerman C B, Miles R D, Littell R C (1997): Relative bioavailability of supplemental inorganic zinc sources for chicks.. Journal of Animal Science, 75, 3195- https://doi.org/10.2527/1997.75123195xShen Hui, Qin Haihong, Guo Junsheng (2008): Cooperation of metallothionein and zinc transporters for regulating zinc homeostasis in human intestinal Caco-2 cells. Nutrition Research, 28, 406-413 https://doi.org/10.1016/j.nutres.2008.02.011Stark Wendelin J. (2011): Nanoparticles in Biological Systems. Angewandte Chemie International Edition, 50, 1242-1258 https://doi.org/10.1002/anie.200906684Xia Tian, Kovochich Michael, Liong Monty, Mädler Lutz, Gilbert Benjamin, Shi Haibin, Yeh Joanne I., Zink Jeffrey I., Nel Andre E. (2008): Comparison of the Mechanism of Toxicity of Zinc Oxide and Cerium Oxide Nanoparticles Based on Dissolution and Oxidative Stress Properties. ACS Nano, 2, 2121-2134 https://doi.org/10.1021/nn800511kYu J., Baek M. (2011): Effects of physicochemical properties of zinc oxide nanoparticles on cellular uptake. Journal of Physics: Conference Series, 34, 301.