MicroRNA-15b participates in diabetic retinopathy in rats through regulating IRS-1 via Wnt/β-catenin pathway

H.-W. Liu, Y. Meng, Y.-B. Ren, P. Sun

Department of Ophthalmology, First Affiliated Hospital of Jiamusi University, Jiamusi, China. 15694548795@163.com

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• Ophthalmology

OBJECTIVE: To explore the role of microRNA-15b in diabetic retinopathy (DR) and its underlying mechanism.

MATERIALS AND METHODS: Diabetic retinopathy rat model was first constructed. Retinal endothelial cells (EC) and retinal pericytes (RP) in DR rats were extracted. The mRNA expression of microRNA-15b in EC and RP cells was detected by qRT-PCR (quantitative Real Time-Polymerase Chain Reaction). Protein expression of insulin receptor substrate 1 (IRS-1) in EC and RP cells was detected by Western blot. After altering microRNA-15b expression by plasmid transfection, cell viability was detected by CCK-8 (cell counting kit-8) assay. Furthermore, the target gene of microRNA-15b was predicted by TargetScan analysis and the binding condition was verified by luciferase reporter gene assay. Finally, rescue experiments were carried out to explore the regulatory effect of microRNA-15b on IRS-1.

RESULTS: MicroRNA-15b was lowly expressed, whereas IRS-1 was highly expressed in EC and RP cells. After overexpression of microRNA-15b, viabilities of EC and RP cells were decreased and β-catenin expression was inhibited. TargetScan predicted that IRS-1 was the downstream gene of microRNA-15b, which was further verified by luciferase reporter gene assay. Rescue experiments indicated that microRNA-15b was capable of regulating IRS-1 via Wnt/β-catenin signaling pathway.

CONCLUSIONS: MicroRNA-15b participates in the development of diabetic retinopathy by targeting IRS-1 via Wnt/β-catenin signaling pathway.

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