1. lncRNA Ttc3-209 Promotes the Apoptosis of Retinal Ganglion Cells in Retinal Ischemia Reperfusion Injury by Targeting the miR-484/Wnt8a Axis.
- Author
-
Zhang R, Feng Y, Lu J, Ge Y, and Li H
- Subjects
- Animals, Blotting, Western, Cells, Cultured, Disease Models, Animal, Electroretinography, Flow Cytometry, Gene Expression Regulation physiology, In Situ Hybridization, Fluorescence, In Situ Nick-End Labeling, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Microscopy, Fluorescence, Real-Time Polymerase Chain Reaction, Reperfusion Injury pathology, Retinal Diseases genetics, Retinal Vessels pathology, Apoptosis, MicroRNAs genetics, RNA, Long Noncoding genetics, Reperfusion Injury genetics, Retinal Diseases pathology, Retinal Ganglion Cells pathology, Ubiquitin-Protein Ligases genetics, Wnt Proteins genetics
- Abstract
Purpose: Apoptosis of the retinal ganglion cells (RGCs) can cause irreversible damage to visual function after retinal ischemia reperfusion injury (RIR). Using a lncRNA chip assay, we selected lncRNA Ttc-209 and characterized its role in RGCs during ischemia reperfusion (I/R)-induced apoptosis., Methods: We created an ischemic model of RGCs by applying Hank's balanced salt solution containing 10 µM antimycin A and 2 µM calcium ionophore for 2 hours. RIR was induced in mice by elevating the intraocular pressure to 120 mm Hg for 1 hour by cannulation of the cornea; this was followed by reperfusion. Real-time quantitative PCR was used to detect the expression levels of long noncoding RNA (lncRNA), microRNA (miRNA), and target gene mRNA. Western blotting, flow cytometry, immunofluorescent staining, and TUNEL assays were performed to detect cell apoptosis. Dual-luciferase reporter assays and FISH were used to identify endogenous competitive RNA (ceRNA) mechanisms that link lncRNAs, miRNAs, and target genes. We also used scotopic electroretinography examinations to evaluate visual function in treated mice., Results: lncRNA Ttc3-209 was significantly upregulated after I/R injury and played a proapoptotic role in RGCs during I/R-induced apoptosis. Mechanistically, lncRNA Ttc3-209 is a ceRNA that competitively binds to miR-484 and upregulates the translation of its target (Wnt8a mRNA), thus promoting apoptosis in RGCs., Conclusions: Reducing the expression of lncRNA Ttc3-209 had a protective effect against apoptosis in RGCs. This may provide a new therapeutic option for the prevention of RGC apoptosis in response to RIR injury.
- Published
- 2021
- Full Text
- View/download PDF