Long non-coding RNA Arid2-IR affects advanced glycation end products-induced human retinal endothelial cell injury by binding to Smad3.

ABSTRACT

PURPOSE: Long non-coding RNAs (lncRNAs) have been reported to play significant roles in the pathogenesis of diabetic retinopathy (DR). The aim of the present study was to investigate the role of lncRNA Arid2-IR in advanced glycation end product (AGE)-induced human retinal endothelial cells (HRECs) injury. MATERIALS AND METHODS: Cell viability was determined by Cell Counting Kit-8 assay following induction with AGEs. The expression of Arid2-IR and Smad3 was detected by reverse transcription-quantitative PCR or western blotting. A luciferase reporter assay was conducted to determine the interaction between Arid2-IR and Smad3. The levels of inflammation-related and oxidative stress-related factors were evaluated by respective kits. The expression of extracellular matrix (ECM)-related and apoptosis-related proteins was detected by western blotting. Immunofluorescence assay was used to detect the level of vascular endothelial growth factor, and flow cytometry was applied to measure the levels of apoptosis. RESULTS: The results revealed that AGE treatment decreased HREC proliferation and upregulated the expression of Arid2-IR and Smad3. The luciferase assay indicated that Smad3 was able to bind to the promoter region sequence of Arid2-IR. Moreover, Arid2-IR silencing reduced inflammation, oxidative stress and ECM production induced by AGEs in HRECs, and Smad3 inhibition further reduced the levels of the aforementioned factors, while Smad3 overexpression exerted the opposite effect. Furthermore, apoptosis of HRECs induced by AGEs was decreased following Arid2-IR silencing, which was further reduced following treatment with Smad3 inhibitor, but was reversed after transfection with Smad3 pcDNA3.1. CONCLUSION: The findings demonstrated that Arid2-IR affects AGE-induced HREC injury by binding to Smad3.