Adenosine Deaminase-2-Induced Hyperpermeability in Human Retinal Vascular Endothelial Cells Is Suppressed by MicroRNA-146b-3p

作者：Samra Yara A; Saleh Heba M; Hussein Khaled A; Elsherbiny Nehal M; Ibrahim Ahmed S; Elmasry Khaled; Fulzele Sadanand; El Shishtawy Mamdouh M; Eissa Laila A; Al Shabrawey Mohamed^*; Liou Gregory I^*

来源：Investigative Ophthalmology & Visual Science, 2017, 58(2): 933.

DOI：10.1167/iovs.16-19782

• adenosine deaminase-2
• miR-146b-3p
• BRB
• TNF-alpha
• IL-6
• ZO-1
• leukocyte adhesion
• diabetic retinopathy

PURPOSE. We recently demonstrated that adenosine deaminase-2 (ADA2) contributes to diabetic retinopathy (DR) via up-regulating the production of inflammatory cytokines in macrophages. Also, microRNA (miR)-146b-3p has the ability to inhibit ADA2. The goal of this study was to investigate the potential role of ADA2 and therapeutic benefit of miR-146b-3p in retinal inflammation and endothelial barrier dysfunction during diabetes.
METHODS. Adenosine deaminase-2 activity was determined by colorimetric method in diabetic human vitreous. Human monocyte cell line U937 was differentiated into macrophages and then treated with amadori glycated albumin (AGA), and conditioned medium (CM) was used to assess the changes in ADA2 activity and TNF-a and IL-6 levels by ELISA. Also, macrophages were transfected with miR-146b-3p before treatment with AGA. Permeability of human retinal endothelial cells (hRECs) was assessed by electric cell-substrate impedance sensing (ECIS) after treatment with macrophage CM. Zonula occludens (ZO)-1 was examined by immunofluorescence in hRECs. Leukocyte adhesion was assessed in hRECs by measuring myeloperoxidase (MPO) activity and intercellular adhesion molecule-1 (ICAM-1) expression.
RESULTS. Adenosine deaminase-2 activity was significantly increased in diabetic human vitreous. ADA2 activity and TNF-a and IL-6 levels were significantly increased in human macrophages by AGA treatment. Amadori glycated albumin-treated macrophage CM significantly increased hREC permeability, disrupted ZO-1 pattern, and increased leukocyte adhesion to hRECs through up-regulating ICAM-1. All these changes were reversed by miR146b- 3p.
CONCLUSIONS. Adenosine deaminase-2 is implicated in breakdown of the blood-retinal barrier (BRB) in DR through macrophages-derived cytokines. Therefore, inhibition of ADA2 by miR146b- 3p might be a useful tool to preserve BRB function in DR.

• 出版日期2017-2