Abstract
Purpose :
Blue-light (BL) from the Light-emitting diodes (LEDs) causes oxidative stress in the retina, resulting in harmful effects on the eye. The major cell types of retina, neural retinal cells and glial cells, play crucial roles in retinal degeneration induced by BL. In this study, we investigated whether crystallins and miR-325-3p are pivotal regulators in neuro-retinal cell exposed to BL.
Methods :
R-28 cells and retina of rats were exposed to wavelength 460 nm and strength 2000 lux using LED chamber, and cells and retinal tissue were collected after incubation. MTT (3-(4,5-Dimethylthiazol-2-yl) assay was performed for analysis of BL-induced retinal cell viability, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was conducted to analyze cell apoptosis of retinal tissue. RT-qPCR and Western immunoblotting were performed to confirm the expressions of RNA and protein, respectively. EGFP-reporter analysis carried out to determine whether miR-325-3p regulates crystallins mRNA by targeting its binding sites.
Results :
The group exposed to blue light showed a decrease in cell viability of R-28 cells compared to the control group. Additionally, the blue light exposure increased apoptotic retinal cells in rats. The mRNA levels of Cryaa and Crybb3 were significantly reduced in the BL-induced retinal degeneration (RD) group, while the level of miR-325-3p increased. Ectopic expression of miR-325-3p down-regulated the expression of EGFP reporters, each containing the 3'UTR of Cryaa and Crybb3, while the control EGFP reporter (EGFP-C1) was not affected. In contrast, inhibition of miR-325-3p up-regulated the expression of both EGFP reporters.
Conclusions :
In this study, we shown that exposure to BL induces apoptotic neuro-retinal cell death in both cell lines and a rat model. Moreover, BL-induced miR-325-3p leads to attenuate expression of Cryaa and Crybb3, playing a protective role in cells. Our results suggest that the molecular axis between miR-325-3p and crystallins (Cryaa and Crybb3) has an essential role in BL-induced retinal degeneration. Therefore, targeting miR-325-3p and crystallins might be able to protect retinal degeneration caused by BL.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.