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Protective effects of NAC and salubrinal on apoptosis of retinal pigment epithelial cells induced by all-trans retinoic acid

Purpose: Accumulation of endogenous all-trans retinoic acid (ATRA) plays a role in the degeneration of photoreceptor cells and retinal pigment epithelium (RPE) cells, contributing to age-related macular degeneration (AMD). This study attempted to investigate the influence of antioxidant N-acetylcysteine (NAC) and selective endoplasmic reticulum stress (ERS) inhibitor salubrinal on apoptosis of ARPE-19 cells induced by ATRA.

Methods: The RPE cell line (ARPE-19) was treated with ATRA, ATRA+NAC, ATRA+salubrinal or ATRA+NAC+salubrinal and the control was untreated. After 24 h of cell culture, the levels of apoptosis, multicaspase and reactive oxygen species (ROS) were detected by flow cytometry. Western blot analysis was employed to detect the expression of vascular endothelial growth factor-A (VEGF-A), C/EBP homologous protein (CHOP) and cleaved caspase-3 in the groups.

Results: The results of flow cytometry showed that NAC and salubrinal decreased the levels of apoptosis, ROS and multicaspase. ATRA increased VEGF-A levels associated with neovascularisation. NAC and salubrinal inhibited an increase in VEGF-A, CHOP and caspase-3 caused by ATRA in ARPE-19 cells.

Conclusions: In ARPE-19 cells, the levels of ROS and ERS can be increased by ATRA, contributing to apoptosis, which can be effectively inhibited by NAC and salubrinal. Thus, ATRA may play an important role in the prevention, diagnosis and treatment of age-related macular degeneration.

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S2923 Salubrinal Salubrinal is a selective inhibitor of eIF2α dephosphorylation and inhibits ER stress-mediated apoptosis with EC50 of ~15 μM in a cell-free assay. (21) (3)

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