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Lixia Lu, Juan Wang, Min Wang, Zongyi Li, Weiye Li, Guo-Tong Xu; Regulatory Effects of Mir365 and Bdnf in Müller Cells Involved in Diabetic Retina. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1150.
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© ARVO (1962-2015); The Authors (2016-present)
To examine the inhibitory role of Brain-derived neurotrophic factor (BDNF) on neuro protection in diabetic retinas, and to study this regulatory mechanism by microRNA (miRNA) at post-transcriptional level in vivo.
miR365 and its target, bdnf, was predicted by bioinformatics analysis and validated by luciferase assay in vitro. The interaction between the two was detected in rat Müller cells (rMC-1 cell line) by overexpressing or inhibiting mir365 level. Glyoxal (AGEs stress) was used to treat rat Müller cells (rMC-1 cell line), and then the mRNA level of bdnf and mir365 were detected. Further, the level of miRNA of interest in diabetic retina was detected by qRT-PCR. Neurosensory retina tissues were collected at different time points from diabetes onset in STZ (Streptozotocin)-induced diabetic rats ex vivo. The differential expression level of bdnf was validated by qRT-PCR and ELISA. In vivo, a virus based anti-mir365 was delivered into diabetic rats and its protection effect was detected.
bdnf is one of the important targets of Mir365, and their interaction was confirmed by in vitro double luciferase activity assay in HEK293 cell and negative regulation in rMC-1. In 2mM glyoxal treated rMC-1(AGEs stress), the expression of mir365 was enhanced and bdnf was decreased after treatment. In diabetic retina, the levels of mir365 increased in 1 week and 2 week diabetes, while bdnf expression level showed significant decreased from 4 week diabetes onset as compared to normal controls. After anti-mir365 treatment in diabetic rats, the apoptosis in neuro retina was attenuated.
In diabetic retinas, as mir365 expression was badly promoted, one of its important targets, bdnf expression level significantly down-regulated from 4 weeks after diabetes onset. Since neuro protection is one of the most important need in clinic diabetic retinopathy patient, our in vivo result that anti-Mir365 treatment in rat can protect neuro retina from apoptosis is somehow meaningful to the treatment of DR. also miRNAs, as a new molecular therapy agent, show its potent application in near future.
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