miR‑29a/b cluster suppresses high glucose‑induced endothelial‑mesenchymal transition in human retinal microvascular endothelial cells by targeting Notch2

Zhang,Jiayu; Zeng,Yue; Chen,Jiawei; Cai,Daqiu; Chen,Chengwei; Zhang,Sifang; Chen,Zhenguo

doi:10.3892/etm.2019.7323

miR‑29a/b cluster suppresses high glucose‑induced endothelial‑mesenchymal transition in human retinal microvascular endothelial cells by targeting Notch2

Authors:
- Jiayu Zhang
- Yue Zeng
- Jiawei Chen
- Daqiu Cai
- Chengwei Chen
- Sifang Zhang
- Zhenguo Chen
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Affiliations: Department of Ophthalmology, The Third Affiliated Hospital of Wenzhou Medical University, Rui'an, Zhejiang 325200, P.R. China
Published online on: February 27, 2019 https://doi.org/10.3892/etm.2019.7323
Pages: 3108-3116
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Several studies have previously reported that endothelial cells contributed to pathological fibrosis in proliferative diabetic retinopathy (PDR) through endothelial‑mesenchymal transition (EndMT); however, the precise mechanism of this interaction has not been completely elucidated. The present study investigated the expression of microRNA (miR)‑29a/b cluster in human retinal microvascular endothelial cells (HRMECs) and examined its functional role in high glucose (HG)‑induced EndMT. HRMECs were exposed to glucose at concentrations of 5, 15, 30 and 50 mM for 7 days and reverse transcription‑quantitative polymerase chain reaction, western blotting and immunofluorescence were conducted to determine the expression of genes associated with miR‑29a/b and EndMT. A luciferase reporter gene assay was also performed to confirm the association between miR‑29a/b and neurogenic locus notch homolog protein 2 (Notch2). The expression levels of miR‑29a/b, and endothelial markers vascular endothelial cadherin and cluster of differentiation 31 were decreased, whereas the expression levels of Notch2 and mesenchymal markers, including α‑smooth muscle actin, fibroblast‑specific protein 1 (also named S100 calcium binding protein A4, S100A4), fibronectin and SNAI1 were increased in HRMECs under HG (30 nM) conditions. In addition, Notch2 was identified as a target of miR‑29a and miR‑29b. Overexpression of miR‑29a/b downregulated the expression of Notch2 and subsequently suppressed HG‑induced EndMT. Taken together, the results of the present study revealed that the miR‑29/Notch2 signaling pathway may participate in the regulation of HG‑induced EndMT, and may serve as a potential molecular target during fibrosis in PDR.

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