Department of Ophthalmology, The Second People's Hospital of Huai'an, Huai'an, China.
Department of Ophthalmology, Huai'an Maternity and Child Health Hospital, Huai'an, China.
J Recept Signal Transduct Res. 2021 Feb;41(1):19-31. doi: 10.1080/10799893.2020.1783556. Epub 2020 Jun 25.
MiR-199a-3p is low expressed in diabetic retinopathy (DR). In the current study, we investigated the effects of miR-199a-3p on DR and the potential mechanisms.
A DR rat model was established, and endothelial cells (ECs) and retinal pericytes (RPs) were extracted from the DR model rats to detect miR-199a-3p expression. Bioinformatics analysis predicted that fibroblast growth factor 7 (FGF7) was a target gene for miR-199a-3p, which was confirmed by dual-luciferase assay. Cell proliferation, migration, and invasion were detected by cell counting kit-8 (CCK-8), colony formation assay, wound-healing, and Transwell assay. Quantitative real-time polymerase chain reaction (q-PCR) and Western blot were performed to detect the expressions of mRNAs and proteins.
MiR-199a-3p was low expressed and FGF7 was high-expressed in ECs and RPs. Overexpressed miR-199a-3p suppressed the proliferation, migration, and invasion, and FGF7 expression of ECs and RPs. However, overexpression of FGF7 effectively eliminated the suppressive effects of miR-199a-3p overexpression malignant behaviors of the cells. Meanwhile, up-regulation of FGF7 noticeably reversed the phosphorylation of phosphoinositide 3-kinase (PI3K) and protein kinase B (AKT) and the expression of epidermal growth factor receptor (EGFR) reduced by miR-199a-3p.
Our findings revealed that in the DR rat model, miR-199a-3p inhibited cell proliferation, migration, and invasion of EC and RP through targeting FGF7 and inhibiting the activation of the EGFR/PI3K/AKT pathway. This study may provide a new direction for the search for the treatment of DR.
miR-199a-3p 在糖尿病视网膜病变（DR）中低表达。在本研究中，我们研究了 miR-199a-3p 对 DR 的影响及其潜在机制。
建立 DR 大鼠模型，从 DR 模型大鼠中提取内皮细胞（EC）和视网膜周细胞（RP），检测 miR-199a-3p 的表达。生物信息学分析预测成纤维细胞生长因子 7（FGF7）是 miR-199a-3p 的靶基因，通过双荧光素酶报告基因实验进行验证。通过细胞计数试剂盒-8（CCK-8）、集落形成实验、划痕实验和 Transwell 实验检测细胞增殖、迁移和侵袭能力。采用定量实时聚合酶链反应（q-PCR）和 Western blot 检测 mRNA 和蛋白的表达。
miR-199a-3p 在 EC 和 RP 中低表达，FGF7 高表达。过表达 miR-199a-3p 抑制了 EC 和 RP 的增殖、迁移和侵袭以及 FGF7 的表达。然而，过表达 FGF7 可有效消除 miR-199a-3p 过表达对细胞恶性行为的抑制作用。同时，上调 FGF7 可明显逆转 miR-199a-3p 下调引起的磷酸肌醇 3-激酶（PI3K）和蛋白激酶 B（AKT）磷酸化以及表皮生长因子受体（EGFR）表达降低。
在 DR 大鼠模型中，miR-199a-3p 通过靶向 FGF7 抑制 EGFR/PI3K/AKT 通路的激活，抑制 EC 和 RP 的增殖、迁移和侵袭。本研究为寻找 DR 的治疗方法提供了新的方向。
