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miR-25 Mediates Retinal Degeneration Via Inhibiting ITGAV and PEDF in Rat

[ Vol. 17 , Issue. 5 ]

Author(s):

J. Zhang, J. Wang, L. Zheng, M. Wang, Y. Lu, Z. Li, C. Lian, S. Mao, X. Hou, S. Li, J. Xu, H. Tian, C. Jin, F. Gao, J. Zhang, F. Wang, W. Li, L. Lu* and G.-T. Xu*   Pages 359 - 374 ( 16 )

Abstract:


Background: Age-related macular degeneration (AMD) is the main cause of irreversible blindness in the elderly. Oxidative stress in retinal pigment epithelium (RPE) is deemed to play a pivotal role in the pathogenesis of AMD. miR-25 functions as an essential modulator in response to oxidative-stress in several cell types, but its function in RPE cells is poorly understood.

Objective: To explore the roles of miR-25 in RPE cells and in the development of AMD.

Methods: A rat model of retinal degeneration was induced by sodium iodate (SI). Subretinal injection of antagomiR-25 was performed for the intervention while the scramble as control. Visual responses were recorded with Electroretinogram (ERG). TUNEL assay was performed to detect apoptosis. Phagosome quantification in vivo was performed to evaluate RPE cell function. Oxygen-glucose deprivation treatment was performed to mimic in vitro oxidative stress. Gene expression at mRNA level and protein level were performed by quantitative polymerase chain reaction (qPCR) and Western Blot, respectively. The pigment epithelium derived factor (PEDF) level in the cultured medium was measured by Enzyme-linked immunosorbent assay (ELISA). The interaction between miR-25 and integrin αV (IGTAV) / PEDF 3’UTR was examined by dual luciferase assay. Chromatin immunoprecipitation (ChIP) assay was performed to examine its transcriptional regulation of miR-25.

Results: Oxidative stress up-regulated miR-25 in RPE cells in very early stage, accompanied by decreased phagocytosis and reduced growth factor secretion in those cells. Such changes preceded RPE cell apoptosis and visual impairment in the SItreated rats. Furthermore, antagomiR-25 intervention effectively rescued RPE cells from degeneration in such model. The increased miR-25 was confirmed to mediate RPE degeneration through direct targeting IGTAV and PEDF. On the other hand, upstream, miR-25 was found to be up-regulated by STAT3 signaling under oxidative stress in both in vivo and in vitro models.

Conclusion: Our findings demonstrate that, in SI-treated rats, oxidative stress activates STAT3 signaling which up-regulates miR-25 expression, in a very early stage. The increased miR-25 then inhibits ITGAV and PEDF expressions, resulting in RPE phagocytosis dysfunction and then RPE apoptosis and visual impairment as observed in patients with AMD. These findings lead us to a better understanding of AMD pathogenesis, and suggest that miR-25 could be a potential therapeutic target for oxidative stress related RPE diseases, like AMD.

Keywords:

Retinal degeneration, miR-25, oxidative-stress, RPE cell, ITGAV, PEDF, STAT3.

Affiliation:

Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Department of Ophthalmology, Drexel University College of Medicine, Philadelphia, PA, Laboratory of Clinical Visual Science, Department of Regenerative Medicine, and Stem Cell Research Center, Tongji University School of Medicine, Shanghai, Department of Ophthalmology of Shanghai Tenth People's Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai



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