Purchase this article with an account.
Juan Wang, Maihemuti Awuti, Xiaoman Jiang, Jieping Zhang, Yali Lyu, Lixia Lu, Guo-Tong Xu; Effect of Glia Maturation Factor beta (GMFB) on Retinal Pigment Epithelium in Diabetic Retinopathy. Invest. Ophthalmol. Vis. Sci. 2018;59(9):3979. doi: https://doi.org/.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To examine the inhibitory role of glia maturation factor beta (GMFB) in diabetic model retinal pigment epithelium (RPE) cell, and to study possible mechanism of Gmfb in diabetic rat RPE.
RPE-Bruch’s membrane-choriocapillaris complex (RBCC) tissues were collected at different time points from diabetes onset in STZ (Streptozotocin)-induced diabetic rats ex vivo. The differential expression level of Gmfb was validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunostaining and Western Blot. Further additional 25mM glucose was used to treat ARPE19 cell line in vitro, and then role of GMFB in RPE cells, including high glucose induced release of inflammatory factors, expression of vascular endothelial growth factor (Vegf), poly ADP-ribose polymerase 1(PARP1) and apoptosis were detected. After treatment with rGMFB in ARPE19 for 24h, RNAseq and bioinformatic analysis were applied.
Gmfb/GMFB mRNA and protein expression level was significantly up-regulated in diabetic rat RBCC from 1w diabetic onset. Immunoreactivity GMFB was up-regulated in diabetic rat RPE cells after 2w diabetic onset as compared to normal controls. In high glucose treated ARPE19 cell, the mRNA and protein expression of Gmfb/GMFB increased, reached peak after 2h treatment and then decreased, with no difference at 24h when compared with normal control. Inflammatory factors IL-1beta, IL-6, IL-33 levels was significantly increased, while VEGF, PARP1 (cleaved) and CASPASE-3(cleaved) protein expression level was significantly up-regulated by ELISA. RNAseq and bioinformatics analysis rGMFB treated ARPE19 elucidated possible signaling pathways including Chemokine signaling pathway, Focal adhesion, ECM-receptor interaction and Wnt signaling pathway.
GMFB expression level was up-regulated in RPE in vivo and in vitro under high glucose condition. GMFB was related with release of inflammatory factors, increased expression of VEGF and increased apoptosis. GMFB knockdown reduce high glucose induced release of inflammatory factors, increased expression of VEGF and increased apoptosis. GMFB related with cell metabolisim, response to stimulus and cellular process.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
This PDF is available to Subscribers Only