Neurotoxicity Research

, Volume 32, Issue 4, pp 624–638 | Cite as

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

  • Marzieh Sharifi Pasandi
  • Farshad Hosseini Shirazi
  • Mohammad Reza Gholami
  • Hossein Salehi
  • Nowruz Najafzadeh
  • Mohammad Mazani
  • Hatef Ghasemi Hamidabadi
  • Ali NiapourEmail author


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.


Epi/perineural cells Schwann cells 2,4-D Apoptosis Claudin-1 ZO-1 



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.

Author 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.

Compliance with Ethical Standards


The authors declare no potential conflicts of interest.


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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Marzieh Sharifi Pasandi
    • 1
    • 2
  • Farshad Hosseini Shirazi
    • 3
    • 4
  • Mohammad Reza Gholami
    • 5
  • Hossein Salehi
    • 6
  • Nowruz Najafzadeh
    • 7
  • Mohammad Mazani
    • 2
  • Hatef Ghasemi Hamidabadi
    • 8
  • Ali Niapour
    • 7
    Email author
  1. 1.Student Research CommitteeArdabil University of Medical SciencesArdabilIran
  2. 2.Department of Biochemistry, School of MedicineArdabil University of Medical SciencesArdabilIran
  3. 3.Department of Toxicology, School of PharmacyShahid Beheshti University of Medical SciencesTehranIran
  4. 4.Pharmaceutical Sciences Research CenterShahid Beheshti University of Medical SciencesTehranIran
  5. 5.Department of Anatomical Sciences, School of MedicineKermanshah University of Medical SciencesKermanshahIran
  6. 6.Department of Anatomical Sciences and Molecular Biology, School of MedicineIsfahan University of Medical SciencesIsfahanIran
  7. 7.Research Laboratory for Embryology and Stem Cells, Department of Anatomical Sciences, School of MedicineArdabil University of Medical SciencesArdabilIran
  8. 8.Immunogenetic Research Center, Department of Anatomy and Cell Biology, School of MedicineMazandaran University of Medical SciencesSariIran

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