miR-205-5p suppresses high glucose-induced angiogenesis in retinal cells

Abstract

Purpose : Retinal angiogenesis and endothelial cell dysfunction are critical complications of Diabetic Retinopathy (DR). miR-205-5p has been pointed out as an anti-angiogenic factor, suppressing angiogenesis in different organs. Therefore, this study tested the hypothesis that miR-205-5p prevents retinal damage under high glucose conditions by regulating HIF-1α and VEGFA.

Methods : The conditioned medium from ARPE-19 cells treated with 35 mM glucose and/or transfected with miR-205-5p mimic and/or treated with 4 mM N-Acetylcysteine (NAC) was exposed to HUVEC cells and angiogenesis was analyzed. The total tube length from images was quantified by Image J using the Angiogenesis Analyzer plugin. VEGFA and HIF-1α mRNA levels from ARPE-19 cells treated under different experimental conditions were evaluated by quantitative real-time PCR (qRT-PCR). Relative expression was analyzed using the 2-ΔΔCt method. Group variation was calculated using a one-way ANOVA with Tukey’s multiple comparison test. Values are expressed as mean±SEM (angiogenesis assay n=3 and qRT-PCR n=4).

Results : HUVEC tube formation was significantly increased by the addition of the medium of ARPE-19 cells treated with 35 mM glucose (202.4±28.7). This effect was normalized by the conditioned medium of ARPE-19 treated with NAC (127.9±12.24), miR-205-5p mimic (111.7±12.57), and a combination of both (113.1±15.53; Figure 1). Overexpression of miR-205-5p led to a decrease in HIF-1α mRNA expression under both control conditions (0.372±0.097) and high-glucose conditions (0.341±0.037). Moreover, it decreased VEGFA mRNA expression exclusively in high-glucose (0.605±0.103) conditions (Figure 2). Additionally, the addition of NAC to the miR-205-5p mimic did not produce additional effects, proposing an absence of synergistic interaction between the two.

Conclusions : miR-205-5p inhibits angiogenesis in the retina by reducing the expression of HIF-1α and VEGFA, suggesting that upregulation of miR-205-5p could serve as an antiangiogenic therapy for DR.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.