Diabetes, Metabolic Syndrome and Obesity (Mar 2021)
Silencing of miR-20b-5p Exerts Inhibitory Effect on Diabetic Retinopathy via Inactivation of THBS1 Gene Induced VEGF/Akt/PI3K Pathway
Abstract
YanBo Ma,1 ChunYing Dong,2 XiHui Chen,1 RuiXi Zhu,1 Jie Wang1 1Department of Ophthalmology, Heilongjiang Provincial Hospital, Harbin, Heilongjiang, 150036, People’s Republic of China; 2Department of Infectious Disease, Heilongjiang Provincial Hospital, Harbin, Heilongjiang, 150036, People’s Republic of ChinaCorrespondence: Jie WangDepartment of Ophthalmology, Heilongjiang Provincial Hospital, Harbin, Heilongjiang, 150036, People’s Republic of ChinaTel/Fax +86-13656838933Email [email protected]: Diabetic retinopathy (DR) is a damaging complication of the eye. Studies investigating molecular mechanisms of DR are lacking, leading to poor clinical outcomes. miR-20b-5p is up-regulated in DR. The present study aimed to confirm the involvement of miR-20b-5p in DR and the mechanism involved.Methods: Microarray analysis was done to study the differentially expressed miRs. DR model was established using Sprague-Dawley rats, the expression of miR-20b-5p was altered using inhibitor or mimic as treatment. THBS1 was one of the potential genes identified by microarray bioinformatics analysis associated with DR. The expression of THBS1 was suppressed by siRNA to study the mechanism behind involvement of miR-20b-5p in DR. In addition, the levels of miR-20b-5p VEGF/PI3K/Akt pathway associated genes were studied. Correlation between THBS1 and miR-20b-5p was evaluated. Cell apoptosis, growth and tube formation assay was performed.Results: The retinal tissues of DR rats showed over-expressed miR-20b-5p and decreased THBS1 via VEGF/PI3K/Akt cascade. THBS1 was confirmed as the target gene of miR-20b-5p by dual-luciferase reporter gene assay. Upregulation of miR-20b-5p or knockdown of THBS1 caused increased tube formation and cell proliferation, whereas it blocked the cell apoptosis of endothelial cells in rats.Conclusion: The outcomes suggested that silencing of miR-20b-5p resulted in inhibition of tube formation and cell growth in vascular endothelial cells of rats subjected to DR altering the VEGF/PI3K/Akt cascade by up-regulation of THBS1.Keywords: diabetic retinopathy, miR-20b-5p, VEGF/PI3K/Akt, endothelial cells
