Müller Cells Harboring Exosomal lncRNA OGRU Modulate Microglia Polarization in Diabetic Retinopathy by Serving as miRNA Sponges

Abstract

Diabetic retinopathy (DR) is one of the most common complications of diabetes mellitus which is associated with visual loss and blindness worldwide. However, the effective treatments for both early- and late-stage DR remains lacking. A streptozotocin (STZ)-induced diabetic mice model and high glucose (HG)-treated Müller cell model were established. M1/M2 microglia polarization was assessed by immunofluorescence (IF) staining and flow cytometry. Expression of lncRNA OGRU, cytokines and other key molecules were detected by qRT-PCR or western blot. ELISA assay was employed to monitor cytokine secretion. Müller cell-derived exosomes were isolated and characterized by nanopartical tracking analysis (NTA), western blot and transmission electron microscopy (TEM), and exosome uptake assay was used to monitor the intercellular transport of exosomes. Associations among lncRNA-miRNA-mRNA networks were validated by RNA pull-down and RNA immunoprecipitation (RIP) and dual luciferase assays. Increased M1 polarization but decreased M2 polarization of retinal microglia were observed in DR mice. HG-treated Müller cell-derived exosomes transported OGRU into microglia and promoted microglia polarization toward M1 phenotype. Mechanistically, OGRU served as a competing endogenous RNA (ceRNA) for miR-320-3p, miR-221-3p and miR-574-5p to regulate AR, PFKFB3 and GLUT1 expression in microglia, respectively. Loss of miR-320-3p/miR-221-3p/miR-574-5p or reinforced AR/PFKFB3/GLUT1 abrogated OGRU silencing-mediated microglia polarization in vitro. In vivo studies further showed that OGRU/miR-320-3p/AR, OGRU/miR-221-3p/PFKFB3 and OGRU/miR-574-5p/GLUT1 axes regulated microglia polarization in DR mice. Collectively, Müller cells-derived exosomal OGRU regulated microglia polarization in DR via modulating OGRU/miR-320-3p/AR, OGRU/miR-221-3p/PFKFB3 and OGRU/miR-574-5p/GLUT1 axes.