Ascorbate Suppresses VEGF Expression in Retinal Pigment Epithelial Cells.

ABSTRACT:

Purpose

To investigate the impact of ascorbate, via DNA hydroxymethylation, on VEGF expression in retinal pigment epithelial (RPE) cells.

Methods

Dot-blot and hydroxymethylated DNA immunoprecipitation sequencing were applied to evaluate the impact of ascorbate on DNA hydroxymethylation in ARPE-19 cells. RNA sequencing (RNA-seq) was carried out to analyze the transcriptome. Quantitative RT-PCR and ELISA were conducted to examine the transcription and secretion of VEGF from cultured cells. Primary human fetal RPE cells and RPE-J cells were used to verify the effect of ascorbate on VEGF expression. ELISA was used to measure VEGF in the vitreous humor of Gulo-/- mice, which, like humans, cannot synthesize ascorbate de novo.

Results

Treatment with ascorbate (50 μM) promote

SUBMITTER: Sant DW 

PROVIDER: S-EPMC6049987 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature