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Epi/perineural and Schwann Cells as Well as Perineural Sheath Integrity are Affected Following 2,4-D Exposure

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Abstract

2,4-dicholorophenoxy acetic acid (2,4-D) is a worldwide-known hormone herbicide. However, there are increasing concerns about its exposure and risks of developing pathological conditions for the peripheral nervous system. The aim of this study was to investigate the mechanism(s) involved in the toxicity of 2,4-D on peripheral nerve’s cellular components. The epi/perineural and Schwann cells and a total of three cell lines were treated with 2,4-D. The viability of cells at different doses of 2,4-D was measured by MTT assay. The cell cycle analyses, cumulative cell counting, fluorescent staining, antioxidant and caspase enzymes activity were examined on epi/perineural and Schwann cells. The epi/perineural cells were assessed as having biological macromolecular changes. Some tight junction-related genes and proteins were also tested on explants of 2,4-D treated epi/perineural tissue. The viability of 2,4-D treated cells was reduced in a dose-dependent manner. Reduced growth rate and G1 cell cycle arrest were verified in 2,4-D treated epi/perineural and Schwann cells. The use of staining methods (acridine orange/ethidium bromide and DAPI) and caspase 3/7 activity assay along with malondialdehyde, glutathione peroxidase, and superoxide dismutase activity assays indicated the apoptotic and oxidant effects of 2,4-D on epi/perineural and Schwann cells. Data obtained from FTIR revealed changes in epi/perineural proteins and cell membrane lipids. Additionally, claudin-1, occludin, and ZO-1 gene/protein expression profiles were significantly reduced in 2,4-D-treated epi/perineural pieces. Our data indicated that oxidative stress, apoptosis of epi/perineural and Schwann cell and impaired blood-nerve barrier may have contributed to nerve damage following 2,4-D exposure.

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Acknowledgements

This study was financially supported by a thesis grant for Master of Science (grant no. 019) and a research project (grant no. 9223) of Research Vice-Chancellery of Ardabil University of Medical Sciences, Iran.

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Authors and Affiliations

  1. Student Research Committee, Ardabil University of Medical Sciences, Ardabil, Iran

    Marzieh Sharifi Pasandi

  2. Department of Biochemistry, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

    Marzieh Sharifi Pasandi & Mohammad Mazani

  3. Department of Toxicology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Farshad Hosseini Shirazi

  4. Pharmaceutical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Farshad Hosseini Shirazi

  5. Department of Anatomical Sciences, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran

    Mohammad Reza Gholami

  6. Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

    Hossein Salehi

  7. Research Laboratory for Embryology and Stem Cells, Department of Anatomical Sciences, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

    Nowruz Najafzadeh & Ali Niapour

  8. Immunogenetic Research Center, Department of Anatomy and Cell Biology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran

    Hatef Ghasemi Hamidabadi

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  1. Marzieh Sharifi Pasandi
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  2. Farshad Hosseini Shirazi
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  3. Mohammad Reza Gholami
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  4. Hossein Salehi
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  7. Hatef Ghasemi Hamidabadi
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  8. Ali Niapour
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Contributions

Cell culture experiments were performed by M.S.P. and A.N. DAPI, EB/AO, staining and caspase assay were photographed and assessed by M.S.P. and A.N. Flow-cytometry was performed and interpreted by A.N. and N.N. FTIR analysis was applied by F.H.S. Biochemical analysis was performed under supervision of M.M. Tissue viability assay, RT-PCR, and western blotting were performed by M.S.P., A.N., and H.S. The paper was written principally by A.N. with input from all the other authors, especially H.S. Study design and statistically analyzing of data were done by A.N., M.R.G., and H.G.H.

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Correspondence to Ali Niapour.

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Sharifi Pasandi, M., Hosseini Shirazi, F., Gholami, M.R. et al. Epi/perineural and Schwann Cells as Well as Perineural Sheath Integrity are Affected Following 2,4-D Exposure. Neurotox Res 32, 624–638 (2017). https://doi.org/10.1007/s12640-017-9777-y

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