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. 2010 Feb 2:4:33-9.
doi: 10.2147/opth.s7979.

Evaluation of ultraviolet light toxicity on cultured retinal pigment epithelial and retinal ganglion cells

Sankarathi Balaiya  1 , Ravi K Murthy, Vikram S Brar, Kakarla V Chalam
Affiliations

Evaluation of ultraviolet light toxicity on cultured retinal pigment epithelial and retinal ganglion cells

Sankarathi Balaiya et al. Clin Ophthalmol. .

Abstract

Purpose: Our study is aimed at evaluating the role of UVB light in inducing cytotoxicity in an in vitro model.

Methods: RGC-5 and ARPE-19 cells were exposed to different time periods of UVB light: 0, 15, 30, and 45 min. They were subsequently examined for changes in cell morphology, cell viability (neutral red uptake assay), generation of reactive oxygen species (ROS), expression of bax, bcl-2 and cytochome C by reverse transcriptase polymerase chain reaction and western blot, respectively.

Results: Dose-dependent reduction in cell viability to UVB light was demonstrated with parallel increase in ROS. Increased duration of exposure (>15 minutes), was associated with increased expression of bax and cytochrome C, and absence of bcl-2 expression.

Conclusion: UVB light exposure results in cell cytotoxicity. The concomitant generation of ROS and expression of apoptotic markers suggests the role of oxidative stress in UVB-mediated apoptosis in an in vitro model of retinal ganglion and pigment epithelial cells.

Keywords: cytochrome C; reactive oxygen species; retinal ganglion cell; retinal pigment epithelium; ultraviolet light.

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Figures

Figure 1
Figure 1
Phase contrast micrographs show the presence of morphological changes such as bleb formation and membrane rupture observed in RGC-5 cells after various time points of UVB exposure.
Figure 2
Figure 2
Phase contrast micrographs shows the presence of morphological changes such as bleb formation, vacuolation, and membrane rupture observed in ARPE-19 cells after various time points of UVB exposure.
Figure 3
Figure 3
Viability of retinal ganglion and pigment epithelial cells after 15, 30, and 45 min exposure to UVB light. Notes: *P < 0.00005, **P < 0.0005.
Figure 4
Figure 4
ROS levels assessed by DHR123 after exposure to different time points of UVB exposure. Note: *P < 0.00005. Abbreviation: ROS, reactive oxygen species.
Figure 5
Figure 5
Electrophoregram illustrating the presence of pro (Bax, 486 bp) and antiapoptotic marker (Bcl-2, 128 bp) expression in ARPE-19 and RGC-5 cells after 0, 15, 30, 45 min exposure to UV radiation. Notes: A, RGC-5; B, ARPE-19; L, 100 bp ladder; Control, GAPDH (348 bp).
See this image and copyright information in PMC

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