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Role of miR-146a in the Regulation of Inflammation in an In Vitro Model of Graves' Orbitopathy

Authors
 Sun Young Jang  ;  Min Kyung Chae  ;  Joon H. Lee  ;  Eun Jig Lee  ;  Jin Sook Yoon 
Citation
 Investigative Ophthalmology & Visual Science, Vol.57(10) : 4027-4034, 2016 
Journal Title
 Investigative Ophthalmology & Visual Science 
ISSN
 0146-0404 
Issue Date
2016
MeSH
Blotting, Western ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Female ; Fibroblasts/metabolism ; Fibroblasts/pathology ; Gene Expression Regulation* ; Graves Ophthalmopathy/genetics* ; Graves Ophthalmopathy/metabolism ; Graves Ophthalmopathy/pathology ; Humans ; Inflammation/genetics ; Inflammation/metabolism* ; Inflammation/pathology ; Interleukin-6/metabolism ; Male ; MicroRNAs/biosynthesis ; MicroRNAs/genetics* ; Microarray Analysis ; RNA/genetics* ; Real-Time Polymerase Chain Reaction
Keywords
miR-146a ; inflammation ; Graves’ orbitopathy
Abstract
PURPOSE: To investigate the role of microRNA 146a (miR-146a) in the regulation of inflammation in an in vitro model of Graves' orbitopathy (GO). METHODS: The level of miR-146a expression in orbital adipose tissue was compared between GO and non-GO by quantitative real-time PCR (qPCR). The effects of interleukin 1β (IL-1β) on miR-146a expression were analyzed in orbital fibroblasts by qPCR. To investigate the molecular mechanism underlying IL-1β-induced miR-146a expression, the effects of inhibitors of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), mitogen-activated protein kinase/extracellular signal-regulated kinases (MEK)-1/2, c-Jun N-terminal kinases (JNK)-1/2, p38 MAP kinase, and phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) were analyzed. The effects of miR-146a mimics and inhibitors on IL-1β-induced IL-6 release were examined by ELISA and Western blotting. RESULTS: The level of miR-146a expression was significantly higher in GO orbital adipose tissue than in non-GO (P = 0.032). Interleukin 1β induced a time- and concentration-dependent increase in miR-146a expression. Interleukin 1β (10 ng/mL, 16 hours) induced an approximately 17.5-fold increase in miR-146 expression. The increase in miR-146a expression by IL-1β was significantly inhibited by NF-κB, JNK-1/2, and PI3K inhibitors (1.94? ± ?0.25, 5.28? ± ?0.34 and 9.73? ± ?2.32-fold, respectively, P < 0.05 compared with IL-1β-induced miR-146 expression, independent t-test). Interleukin 1β-induced IL-6 protein production was further decreased by miR-146a mimics, but not by inhibitors of miR-146a. CONCLUSIONS: MicroRNA 146a was upregulated by inflammatory stress in orbital fibroblasts. Our results indicated that miR-146a had a positive effect on the anti-inflammatory process. MicroRNA 146a may play a role in the regulation of inflammation in orbital fibroblasts, and may participate in the pathogenesis of GO.
Full Text
http://iovs.arvojournals.org/article.aspx?articleid=2543210
DOI
10.1167/iovs.16-19213
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Ophthalmology (안과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
Yonsei Authors
윤진숙(Yoon, Jin Sook) ORCID logo https://orcid.org/0000-0002-8751-9467
이은직(Lee, Eun Jig) ORCID logo https://orcid.org/0000-0002-9876-8370
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URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/151927
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