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The effect of clonidine on VEGF expression in human retinal pigment epithelial cells (ARPE-19)

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Abstract

Background

The purpose of this study was to investigate the effect of clonidine, an alpha2-adrenergic receptor (α2-ADR) agonist, on vascular endothelial growth factor (VEGF) expression and secretion in the human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin-1β (IL-1β).

Methods

Alpha2-ADRs (α2A, α2B, and α2C) mRNA expression in ARPE-19 cells was examined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Clonidine and inhibitors against protein kinases that are involved in the regulation of the intracellular signal transduction were added to serum-free medium before stimulation of IL-1β. The α2-ADR antagonist, Yohimbine, was loaded 30 min before the addition of clonidine. The expression of VEGF mRNA and protein was measured by real-time PCR and enzyme-linked immunosorbent assay.

Results

Alpha2A-ADR, α2B-ADR, and α2C-ADR mRNA was expressed in RPE cells. Clonidine, an inhibitor of p38MAPK and MEK1/2, inhibited the expression of VEGF protein and mRNA in the RPE cells stimulated with IL-1β. The inhibitory effect of clonidine on the secretion of VEGF protein stimulated with IL-1β was blocked by α2-ADR antagonists.

Conclusions

The effect of clonidine on the expression of VEGF may be via suppression of the p38MAPK and MEK1/2 signal transduction pathways activated with IL-1β.

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Acknowledgments

This work was supported in part by a Grant-in-Aid for Scientific Research (No.: 19791259) from the Ministry of Education, Science, Sports, and Culture of Japan.

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Authors and Affiliations

  1. Department of Ophthalmology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan

    Kazuhiko Watanabe, Xue-Yun Zhang, Kiyotaka Kitagawa, Tatsuya Yunoki & Atsushi Hayashi

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  1. Kazuhiko Watanabe
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  2. Xue-Yun Zhang
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Correspondence to Atsushi Hayashi.

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The authors have no financial relationship with any organization.

The authors have full control of all primary data, and agree to allow Graefe’s Archive for Clinic and Experimental Ophthalmology to review the data upon request.

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Watanabe, K., Zhang, XY., Kitagawa, K. et al. The effect of clonidine on VEGF expression in human retinal pigment epithelial cells (ARPE-19). Graefes Arch Clin Exp Ophthalmol 247, 207–213 (2009). https://doi.org/10.1007/s00417-008-0990-5

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  • DOI: https://doi.org/10.1007/s00417-008-0990-5

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