Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

MiR-10b alleviates high glucose-induced human retinal endothelial cell injury by regulating TIAM1 signaling

Yaohua Chen¹, Yanqing Zhu¹, Sheng Zhao^2,3

¹Department of Ophthalmology, Nantong Hospital of Traditional Chinese Medicine, Nantong City, Jiangsu Province 226001; ²Department of Ophthalmology, Hwa Mei Hospital, University of Chinese Academy of Sciences; ³Ningbo Institute of Life and Health Industry, University of Chinese Academy of Sciences, Ningbo City, Zhejiang Province 315000, China.

For correspondence:- Sheng Zhao Email: FGYUk81@163.com Tel:+8657483870541

Accepted: 1 August 2020 Published: 31 August 2020

Citation: Chen Y, Zhu Y, Zhao S. MiR-10b alleviates high glucose-induced human retinal endothelial cell injury by regulating TIAM1 signaling. Trop J Pharm Res 2020; 19(8):1577-1583 doi: 10.4314/tjpr.v19i8.2

© 2020 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To investigate the effects of microRNA (miR)-10b on high glucose (HG)-induced human retinal endothelial cell (HREC) injury and the mechanisms involved.

Methods: Levels of miR-10b were measured in HRECs using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) after the addition of glucose (5.5 and 30 mM). Cell viability was measured using Cell Counting Kit-8 assay, while levels of reactive oxygen species (ROS) were determined using fluorimetry. An enzyme-linked immunosorbent assay (ELISA) was used to measure cellular apoptosis. Luciferase reporter assay was used to validate the miR-10b-binding sites of target genes. The levels of T-cell lymphoma invasion and metastasis (TIAM1) and NADPH oxidase-2 (NOX2) were determined using qRT-PCR. Ras-related C3 botulinum toxin substrate 1 (Rac1) activation was evaluated using a pull-down assay. The protein levels of TIAM1 and Rac1 were assayed by western blotting.

Results: After HG stimulation, miR-10b expression was downregulated. Viability of HRECs decreased, whereas ROS production increased. However, the overexpression of miR-10b inhibited apoptosis and ROS production in HG-treated HRECs (p < 0.05), while luciferase reporter analysis revealed a possible binding site for miR-10b to target the 3'-untranslated region (UTR) of TIAM1. In addition, the overexpression of miR-10b distinctly reduced the expression levels of TIAM1 and NOX2, but decreased the activation of Rac1 in HG-treated HRECs (p < 0.05); these inhibitory effects of miR-10b were significantly reversed after TIAM1 application.

Conclusion: MiR-10b alleviates HG-induced HREC injury by regulating TIAM1 signaling. MiR-10b therapy is a potential therapeutic strategy for patients suffering from diabetic retinopathy.

Keywords: MicroRNA-10b, Human retinal endothelial cells, High glucose, TIAM1-Rac1 axis