Wen Min, Wu Rong, Xie Jin-Feng, Chen Xiao-Mei, Wen Nian-Lian, Huang Sheng
School of Medicine, Zunyi Medical University, Zunyi, Guizhou, China.
School of Medicine, Jishou University, Jishou, Hunan, China.
Ann Clin Lab Sci. 2025 Jan;55(1):19-27.
Diabetic retinopathy (DR) is a retinal microangiopathy caused by diabetes mellitus. miRNAs have been shown to be involved in DR-associated micro vessel formation, influence the DR progression, and may be a therapeutic strategy for DR. miRNA-19a-3p is highly expressed in the vitreous of DR patients, however, its mechanism in the occurrence and development of DR is unclear. Therefore, the purpose of this study was to explore the potential mechanism of miRNA-19a-3p in the occurrence and development of DR.
Human retinal microvascular endothelial cells (hRMECs) were treated with 30 mM high glucose for 48 h to construct a DR cell model. Quantitative real-time polymerase chain reaction was performed to detect the expression levels of miR-19a-3p and SOCS1; the cell viability was measured by Cell Counting Kit-8 after the interference or overexpression of miR-19a-3p. Cell scratch assay was used to verify the cell migration ability; tube formation assay to measure the tube formation ability. The targeting relationship between miR-19a-3p and SOCS1 was demonstrated by dual-luciferase reporter gene assay. Western blot was adopted to determine the protein expression levels of FGF2, VEGFA, SOCS1, JAK2, p-JAK2, STAT3, and p-STAT3.
In the hRMECs induced by high glucose, miRNA-19a-3p was increased, while the expression level of SOCS1 was significantly decreased. The cell viability, migration, and tube formation could be markedly inhibited by suppressing the miR-19a-3p expression in hRMECs under high glucose conditions. Besides, the expression level of SOCS1 was remarkably elevated after the inhibition of miR-19a-3p expression. In addition, miR-19a-3p could directly target the negatively regulated SOCS1. The inhibition of SOCS1 expression reversed the effects of miR-19a-3p down-regulation on the viability, migration, and tube formation of hRMECs under high glucose conditions. miR-19a-3p could activate the JAK2/STAT3 signaling pathway by inhibiting the expression of SOCS1, thereby promoting the tube formation.
miRNA-19a-3p can target SOCS1 to regulate JAK2/STAT3/VEGF, thereby exacerbating tube formation in DR. The results of this study provide a new target for the treatment of DR.
糖尿病视网膜病变（DR）是由糖尿病引起的视网膜微血管病变。微小RNA（miRNA）已被证明参与DR相关微血管形成，影响DR进展，可能是DR的一种治疗策略。miRNA-19a-3p在DR患者玻璃体中高表达，然而，其在DR发生发展中的机制尚不清楚。因此，本研究旨在探讨miRNA-19a-3p在DR发生发展中的潜在机制。
用30 mM高糖处理人视网膜微血管内皮细胞（hRMECs）48小时以构建DR细胞模型。采用定量实时聚合酶链反应检测miR-19a-3p和SOCS1的表达水平；在干扰或过表达miR-19a-3p后，用细胞计数试剂盒-8检测细胞活力。采用细胞划痕试验验证细胞迁移能力；采用管腔形成试验测量管腔形成能力。通过双荧光素酶报告基因试验证实miR-19a-3p与SOCS1之间的靶向关系。采用蛋白质免疫印迹法测定FGF2、VEGFA、SOCS1、JAK2、p-JAK2、STAT3和p-STAT3的蛋白表达水平。
在高糖诱导的hRMECs中，miRNA-19a-3p升高，而SOCS1表达水平显著降低。在高糖条件下，抑制hRMECs中miR-19a-3p表达可显著抑制细胞活力、迁移和管腔形成。此外，抑制miR-19a-3p表达后，SOCS1表达水平显著升高。此外，miR-19a-3p可直接靶向负调控的SOCS1。抑制SOCS1表达可逆转miR-19a-3p下调对高糖条件下hRMECs活力、迁移和管腔形成的影响。miR-19a-3p可通过抑制SOCS1表达激活JAK2/STAT3信号通路，从而促进管腔形成。
miRNA-19a-3p可靶向SOCS1调节JAK2/STAT3/VEGF，从而加剧DR中的管腔形成。本研究结果为DR治疗提供了新靶点。
