J Korean Ophthalmol Soc.  2009 Apr;50(4):594-602. 10.3341/jkos.2009.50.4.594.

Effect of Triamcinolone on Angiogenesis-related Factors of Cultured Retinal Pigment Epithelial Cells

• Lee YC
• Yoon TJ
• Choi GJ
• Kim DH

• KMID: 2212262
• DOI: http://doi.org/10.3341/jkos.2009.50.4.594

Abstract

PURPOSE: To examine the effects of triamcinolone on angiogenesis related factors in cultured human retinal pigment epithelial cells.
METHODS
Human retinal pigment epithelial cells were exposed to triamcinolone, cultured in a hypoxic environment, and expression and production of VEGF and PEDF were subsequently tested by RT-PCR and Western blot. Angiogenesis was measured via a tube formation assay using ECV 304 cells and with a migration assay using human dermal microvascular endothelial cells.
RESULTS
Expression and production of VEGF and PEDF were tested by RT-PCR and Western blot, respectively. VEGF abundance was reduced while that of PEDF was unchanged in triamcinolone exposed retinal pigment epithelial cells cultured in hypoxic environment compared with cells with no treatment in hypoxic environment (p<0.05). Tube formation and cell migration were reduced by triamcinolone (p<0.05).
CONCLUSIONS
These results suggest that triamcinolone affects the secretion of angiogenesis-related factors and suppresses neovascularization.

Keyword

Angiogenesis; PDEF; Triamcinolone; VEGF

MeSH Terms

Blotting, Western
Cell Movement
Endothelial Cells
Epithelial Cells
Humans
Retinaldehyde
Triamcinolone
Vascular Endothelial Growth Factor A
Retinaldehyde
Triamcinolone
Vascular Endothelial Growth Factor A

Figure

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  Figure 1. The viablity of ARPE19 cells on exposure to triamcinolone acetonide (TA). Cells viabilty is fair at lower than 100 μM TA.

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  Figure 2. RT-PCR of VEGF and PEDF genes and the densitometric analysis in the ARPE-19 cells to the exposure of triamcinolone acetonide (TA). Quantitative analysis shows that expression of VEGF gene in these cells decreased after exposure to TA (A, B)(p<0.05). However, PEDF gene is not changed (C, D). VEGF=vascular endothelial growth factor; PEDF=pigment epithelium-derived factor.

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  Figure 3. Western blot on VEGF and PEDF production and the densitometric analysis in the ARPE-19 cells to the exposure of triamcinolone acetonide (TA). Quantitative analysis shows that production of VEGF protein in those cells decreased after exposure to TA (A, B)(p<0.05). However, PEDF production did not change (C, D). VEGF=vascular endothelial growth factor; PEDF=pigment epithelium derived factor.

• 

  Figure 4. Inhibition of tube formation by Triamcinolone acetonide (TA). The cells were cultured with the supernatant of HRPE cells and ECV-304 cell medium (1:1). After 6-hour culture, tube-like structure were analyzed. The supernatants of ARPE-19 cells used were collected from the APRE-19 conditioned media exposed to TA. With exposure to TA on ARPE19 cells, tube-like structures of ECV-304 cells decreased as shown in (A-D)(p<0.05).

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  Figure 5. Inhibition of cell migration by Triamcinolone acetonide (TA). Effect of the RPE-conditioned medium on human dermal microvascular endothelial cell (HDMEC) in migration assay. Conditioned medium was harvested from the culture dishes after ARPE 19 cells were exposed to TA for 6 hours. After initial wound scraping, cells were cultured for 24 hours and then observed. The degree of migration activity of HDMECs increased according to exposure to TA concentration (A-D)(p<0.05).

Reference

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