April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Pparß Antagonist Mediates Increases in Human Corneal Epithelial Cell Migration Through P38 Mapk Activation
Author Affiliations & Notes
  • Z. Zhang
    Department of Biological Sciences, SUNY State College of Optometry, New York, New York
  • Z. Pan
    Department of Biological Sciences, SUNY State College of Optometry, New York, New York
  • F. Zhang
    Department of Biological Sciences, SUNY State College of Optometry, New York, New York
  • P. S. Reinach
    Department of Biological Sciences, SUNY State College of Optometry, New York, New York
  • Footnotes
    Commercial Relationships  Z. Zhang, None; Z. Pan, None; F. Zhang, None; P.S. Reinach, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 1973. doi:
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      Z. Zhang, Z. Pan, F. Zhang, P. S. Reinach; Pparß Antagonist Mediates Increases in Human Corneal Epithelial Cell Migration Through P38 Mapk Activation. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1973.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Peroxisome proliferator-activated receptor beta (PPARß) modulation contributes to the control of wound healing in a variety of tissues. In human corneal epithelial cells (HCECs), its role in mediating this response is unknown. Accordingly, we determined if PPARß inhibition with GSK0660, a described antagonist, alters HCEC cell migration using a scratch wound assay.

Methods: : SV40-HCEC grown to 80% confluence were serum-deprived overnight in the presence of 2.5 mM hydroxyurea. A scratch wound assay was performed to characterize the contribution of cell migration to wound closure. Following wounding, closure was monitored in serum-free medium in the continued presence and absence of compounds of interest. The increases in migration elicited by 5ng/ml EGF served as a positive control. The remaining wound areas were photographed at 0, 5 and 24 h and digitized by SigmaScan Pro 5. The CellTiter-Glo luminescent viability assay assessed for cytotoxicity of GSK0660. Immunocytochemistry was employed to identify the expression of PPARß. All experiments were performed in triplicate.

Results: : Expression of PPARß was identified in the nucleus of HCECs. Up to 200 µM GSK0660 was not cytotoxic. With 1 µM GSK0660, the remaining wound area after 24 h decreased to 37 ± 15 % whereas in the untreated control to 73 ± 2 % of the initial wounds. This stimulation was the same as that obtained with EGF (i.e. 39 ± 8 %). Meanwhile, following p38 mitogen activated protein kinase (MAPK) inhibition with 10 µM SB203580 the wound was 77 ± 13 % of the initial wounds. The inhibitory effects of SB203580 on EGF and GSK0660-promoted migration were indistinguishable from one another, leaving open areas of 85 ± 4 % and 84 ± 9 % of the initial wounds, respectively. On the other hand, a PPAR ß agonist, GW0742, had no effect on migration from 25nM to 1µM.

Conclusions: : GSK0660 stimulation of HCECs migration includes stimulation of p38 MAPK since suppression of p38 MAPK activation by SB203580 obviated the effect of GSK0660 on this response. Even though GSK0660 may block PPARß activation, this compound also stimulates p38 MAPK activation. It is unclear if p38 MAPK is upstream or downstream from PPARß in the signaling cascade mediating this response.

Keywords: cornea: epithelium • wound healing 
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