Effects of microRNA‑217 on high glucose‑induced inflammation and apoptosis of human retinal pigment epithelial cells (ARPE‑19) and its underlying mechanism.

Abstract

Diabetic retinopathy is a major complication of diabetes. Increasing evidence has indicated that microRNAs (miRs) serves an important role in diabetic retinopathy. However, the expression and mechanism of miR-217 in high glucose-induced human retinal pigment epithelial cells ARPE-19 is still unclear. Therefore, the aim of this study was to investigate the role of miR-217 in high glucose-induced retinal epithelial cell damage, and further to explore the molecular mechanisms. In our study, we found that compared with control group, miR-217 was upregulated in high glucose-induced ARPE-19 cells. In addition, TargetScan and a dual-luciferase reporter gene assay showed that Sirtuin 1 (SIRT1) was a direct target of miR-217. Then, we performed reverse transcription-quantitative polymerase chain reaction assay and western blot assay to explore the expression of SIRT1 in high glucose-induced ARPE-19 cells. Our results demonstrated that SIRT1 was downregulated at the mRNA and protein levels in high glucose-induced ARPE-19 cells. Then, ARPE-19 cells were transfected with inhibitor control, miR-217 inhibitor or miR-217 inhibitor + SIRT1-small interfering RNA for 6 h, and then the cells were treated with 50 mM D-glucose for 24 h. We then investigated the effects of miR-217 inhibitor on ARPE-19 cell viability and apoptosis. An MTT assay revealed that miR-217 inhibitor significantly increased the viability and decreased the apoptosis of high glucose-induced ARPE-19 cells. ELISA indicated that miR-217 inhibitor significantly reduced the expression of inflammatory factors, such as interleukin (IL)-1β, tumor necrosis factor-α, and IL-6 in high glucose-treated ARPE-19 cells. Additionally, a western blot assay demonstrated that miR-217 inhibitor suppressed the expression of p-p65. The effects of miR-217 inhibitor on high glucose-treated ARPE-19 cells were significantly reversed by the silencing the SIRT1 gene. Therefore, our findings suggested that miR-217 inhibitor protected against retinal epithelial cell damage caused by high glucose via targeting SIRT1, thereby playing a protective role in diabetic retinopathy. Targeting miR-217 may have therapeutic potential in the treatment of diabetic retinopathy.