Xie Shuying, Liu Zehong, Luo Ting, Chen Yifa, Zeng Liqun, Li Xiaoyan
Department of Endocrinology, Ganzhou People's Hospital, No.17 Hongqi Avenue, Ganzhou City, Jiangxi, 341000, China.
In Vitro Cell Dev Biol Anim. 2025 Jul 1. doi: 10.1007/s11626-025-01056-9.
This study explores the mechanism of lncRNA TDRG1 in high glucose (HG)-induced human retinal microvascular endothelial cell (hRMEC) injury. hRMECs were cultured in HG medium, followed by the detection of cell viability, proliferation, migration, and angiogenesis using CCK-8, EdU, Transwell, and tube formation assays. LncRNA TDRG1, miR-7-5p, G3BP2, VEGFA, and CD31 expression in hRMECs was detected by RT-qPCR or western blot. After transfection with lncRNA TDRG1 siRNA or miR-7-5p inhibitor or G3BP2 pcDNA3.1, hRMEC injury induced by HG was evaluated. Dual luciferase, RIP, or RNA pull-down assays were performed to verify the binding of lncRNA TDRG1, miR-7-5p, and G3BP2. HG treatment notably elevated the expressions of lncRNA TDRG1 and G3BP2 in hRMECs but diminished the expression of miR-7-5p. Low expression of lncRNA TDRG1 restrained the proliferation, migration, and angiogenesis of hRMECs while diminishing VEGFA and CD31 expression. Mechanistically, lncRNA TDRG1 upregulated the transcription level of G3BP2 by competitively binding to miR-7-5p. Low expression of miR-7-5p or overexpression of G3BP2 weakened the inhibitory effect of lncRNA TDRG1 silencing on HG-induced hRMEC injury. In conclusion, lncRNA TDRG1 upregulates the transcription level of G3BP2 by competitively binding to miR-7-5p, thus exacerbating HG-induced hRMEC injury.
本研究探讨长链非编码RNA TDRG1在高糖（HG）诱导的人视网膜微血管内皮细胞（hRMEC）损伤中的作用机制。将hRMECs培养于HG培养基中，随后采用CCK-8、EdU、Transwell和管腔形成实验检测细胞活力、增殖、迁移和血管生成。通过RT-qPCR或蛋白质免疫印迹法检测hRMECs中lncRNA TDRG1、miR-7-5p、G3BP2、VEGFA和CD31的表达。转染lncRNA TDRG1 siRNA、miR-7-5p抑制剂或G3BP2 pcDNA3.1后，评估HG诱导的hRMEC损伤。进行双荧光素酶报告基因、RNA免疫沉淀（RIP）或RNA下拉实验以验证lncRNA TDRG1、miR-7-5p和G3BP2之间的结合。HG处理显著提高了hRMECs中lncRNA TDRG1和G3BP2的表达，但降低了miR-7-5p的表达。lncRNA TDRG1低表达抑制了hRMECs的增殖、迁移和血管生成，同时降低了VEGFA和CD31的表达。机制上，lncRNA TDRG1通过竞争性结合miR-7-5p上调G3BP2的转录水平。miR-7-5p低表达或G3BP2过表达减弱了lncRNA TDRG1沉默对HG诱导的hRMEC损伤的抑制作用。总之，lncRNA TDRG1通过竞争性结合miR-7-5p上调G3BP2的转录水平，从而加剧HG诱导的hRMEC损伤。
