Department of Ophthalmology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.
Department of Ophthalmology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.
J Diabetes Complications. 2020 Oct;34(10):107668. doi: 10.1016/j.jdiacomp.2020.107668. Epub 2020 Jul 2.
The apoptosis of human retinal pigment epithelial cells (RPEs) plays a critical role in the pathogenesis of diabetic retinopathy (DR), but the molecular mechanisms remain unclear. In this study, we explored the function of miR-203a-3p in CoCl-induced RPEs apoptosis.
The cellular localization of miR-203a-3p was assessed by in situ hybridization. Luciferase reporter assays were performed to validate that suppressor of cytokine signaling 3(SOCS3) as a direct target of miR-203a-3p. Effects of miR-203a-3p manipulation on RPEs apoptosis were evaluated using TdT-mediated dUTP Nick-End Labeling (TUNEL) and Flow Cytometry. Expression levels of miR-203a-3p was analyzed by RT-PCR, the expression of target proteins was detected by western blot.
miR-203a-3p was found to be located in the RPE layer of the retinas from normal and diabetic rats and SOCS3 was a direct target of miR-203a-3p. miR-203a-3p mimics resulted in improved CoCl-induced apoptosis of RPEs, overexpression of SOCS3 or c-Jun N-terminal kinase (JNK) inhibitor SP600125 reversed the pro-apoptotic effect of miR-203a-3p, to a certain extent.
Our data implied a crucial role of miR-203a-3p as a novel regulator of CoCl-induced RPEs apoptosis through SOCS3. Deregulation of miR-203a-3p/SOCS3/JNK/c-Jun cascade thus may serve as an important contributor to RPEs apoptosis in DR.
人视网膜色素上皮细胞（RPE）的细胞凋亡在糖尿病视网膜病变（DR）的发病机制中起关键作用，但分子机制尚不清楚。在这项研究中，我们探讨了 miR-203a-3p 在 CoCl 诱导的 RPE 细胞凋亡中的作用。
通过原位杂交评估 miR-203a-3p 的细胞定位。荧光素酶报告基因实验验证了 SOCS3 是 miR-203a-3p 的直接靶标。使用 TdT 介导的 dUTP 缺口末端标记（TUNEL）和流式细胞术评估 miR-203a-3p 操作对 RPE 细胞凋亡的影响。通过 RT-PCR 分析 miR-203a-3p 的表达水平，通过 Western blot 检测靶蛋白的表达。
发现 miR-203a-3p 位于正常和糖尿病大鼠视网膜的 RPE 层中，SOCS3 是 miR-203a-3p 的直接靶标。miR-203a-3p 模拟物导致 CoCl 诱导的 RPE 细胞凋亡改善，SOCS3 过表达或 c-Jun N 末端激酶（JNK）抑制剂 SP600125 在一定程度上逆转了 miR-203a-3p 的促凋亡作用。
我们的数据表明，miR-203a-3p 通过 SOCS3 作为 CoCl 诱导的 RPE 细胞凋亡的新型调节因子发挥重要作用。miR-203a-3p/SOCS3/JNK/c-Jun 级联的失调因此可能是 DR 中 RPE 细胞凋亡的重要贡献者。
