Department of Ophthalmology, Shengjing Hospital, China Medical University, Shenyang, China.
Department of Ophthalmology, The Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu, China.
Biochem Biophys Res Commun. 2018 Nov 10;505(4):1236-1243. doi: 10.1016/j.bbrc.2018.10.052. Epub 2018 Oct 15.
Diabetic retinopathy (DR) is a progressive microvascular complication associated with diabetes, and remains the leading cause of preventable blindness worldwide. Recent studies have revealed that microRNAs (miRNAs) were involving in the physiological and pathophysiological processes of diabetes and its microvascular and macrovascular complications. The purpose of the current investigation is to identify the candidate miR-211 as a novel biomarker for occurrence and progression of DR in clinical study and experimental research. Firstly, miR-211 was considered as a candidate miRNA identifying by miRNA microarray analysis, Venn diagram analysis, real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and receiver operating characteristic curve in clinical study. Then, the predicted Sirtuin 1 (SIRT1) may be the target gene of miR-211 searching by TargetScan 7.2. Moreover, miR-211 was significantly up-regulated, while SIRT1 mRNA significantly down-regulated measuring by qRT-PCR, meanwhile, SIRT1 protein was significantly down-regulated in coincidence with SIRT1 mRNA detecting by western blot, and even aggravated associated with diabetes duration in diabetic retinal tissues of vivo experiment. Additionally, miR-211 was directly targeted SIRT1 confirming by dual-luciferase reporter assay. Furthermore, with transfection of antagomiR-211, the apoptosis of HUVECs was significantly suppressed employing by flow cytometry analysis, nevertheless the viability of HUVECs was significantly promoted exploiting by Cell Counting Kit-8 assay. Finally, SIRT1 mRNA and SIRT1 protein were significantly up-regulated testing by qRT-PCR and western blot respectively in hyperglycemic HUVECs transfected with antagomiR-211 of vitro experiment. Consequently, the current clinical study and experimental research imply that serum miR-211 as a novel biomarker with high sensitivity and specificity could be associated with occurrence and progression of DR via targeting SIRT1.
糖尿病视网膜病变（DR）是一种与糖尿病相关的进行性微血管并发症，仍是全球可预防失明的主要原因。最近的研究表明，microRNAs（miRNAs）参与了糖尿病及其微血管和大血管并发症的生理和病理生理过程。本研究旨在通过临床研究和实验研究，确定候选 miR-211 作为 DR 发生和进展的新型生物标志物。首先，通过 miRNA 微阵列分析、Venn 图分析、实时定量逆转录聚合酶链反应（qRT-PCR）和受试者工作特征曲线，在临床研究中认为 miR-211 是候选 miRNA。然后，通过 TargetScan 7.2 预测 Sirtuin 1（SIRT1）可能是 miR-211 的靶基因。此外，通过 qRT-PCR 检测，miR-211 显著上调，而 SIRT1 mRNA 显著下调，同时，通过 Western blot 检测 SIRT1 蛋白与 SIRT1 mRNA 一致下调，甚至在糖尿病视网膜组织中随糖尿病病程加重。此外，通过双荧光素酶报告基因实验证实 miR-211 可直接靶向 SIRT1。此外，通过流式细胞术分析，转染 antagomiR-211 可显著抑制 HUVECs 的凋亡，而通过 Cell Counting Kit-8 测定法可显著促进 HUVECs 的活力。最后，通过 qRT-PCR 和 Western blot 分别检测转染 antagomiR-211 的高糖 HUVECs 中 SIRT1 mRNA 和 SIRT1 蛋白的表达，发现 SIRT1 mRNA 和 SIRT1 蛋白均显著上调。因此，本临床研究和实验研究表明，血清 miR-211 作为一种新型生物标志物，具有较高的敏感性和特异性，可能通过靶向 SIRT1 与 DR 的发生和进展有关。
