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

Balaiya S, Murthy RK, Brar VS, Chalam KV - Clin Ophthalmol (2010)

Bottom Line: 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.Dose-dependent reduction in cell viability to UVB light was demonstrated with parallel increase in ROS.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, University of Florida College of Medicine, Jacksonville, FL, USA.

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.

No MeSH data available.


Related in: MedlinePlus

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).
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f5-opth-4-033: 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).

Mentions: Apoptotic and antiapoptotic markers, Bax (486 bp) and Bcl-2 (128 bp) were confirmed against 1 kb ladder in 2% and 3% agarose gel electrophoresis, respectively. No bands were detected using apoptotic bax primer in experimental cells up to 15 min after UV treatment. Cells harvested after 30 to 45 min exposure showed a significant increase of bax and a decrease or no band in bcl-2 relative to control. cDNA levels were normalized using house-keeping gene GAPDH (Figure 5).


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

Balaiya S, Murthy RK, Brar VS, Chalam KV - Clin Ophthalmol (2010)

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).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2819767&req=5

f5-opth-4-033: 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).
Mentions: Apoptotic and antiapoptotic markers, Bax (486 bp) and Bcl-2 (128 bp) were confirmed against 1 kb ladder in 2% and 3% agarose gel electrophoresis, respectively. No bands were detected using apoptotic bax primer in experimental cells up to 15 min after UV treatment. Cells harvested after 30 to 45 min exposure showed a significant increase of bax and a decrease or no band in bcl-2 relative to control. cDNA levels were normalized using house-keeping gene GAPDH (Figure 5).

Bottom Line: 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.Dose-dependent reduction in cell viability to UVB light was demonstrated with parallel increase in ROS.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, University of Florida College of Medicine, Jacksonville, FL, USA.

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.

No MeSH data available.


Related in: MedlinePlus