高糖条件下miR-615-5p对人视网膜内皮细胞的影响研究

目的 探讨高糖条件下miR-615-5p对人视网膜内皮细胞(hRECs)迁移能力、成管能力、愈合能力的影响及其机制.方法 取对数生长期的hRECs,将细胞分为低糖(LG)组、高糖(HG)组、miR-615-5p模拟物(mimic)组、miR-615-5p模拟物阴性对照(mi-nc)组、miR-615-5p抑制剂(inh...

Effect of miR-615-5p on human retinal endothelial cells under high glucose conditions

Objective　To investigate the effect and mechanism of miR-615-5p on migration, tube formation, and wound healing of human retinal endothelial cells (hRECs) under high glucose conditions.Methods　hRECs in the logarithmic growth phase were divided into the low glucose (LG) group, high glucose (HG) group, miR-615-5p mimic (mimic) group, miR-615-5p mimic negative control (mi-nc) group, miR-615-5p inhibitor (inhibitor) group, and miR-615-5p inhibitor negative control (in-nc) group. Cells in the LG group were cultured in the medium containing 5.5 mmol·L^-1 glucose, and cells in the other groups were cultured in the medium containing 25.0 mmol·L^-1 glucose. The relative expression levels of miR-615-5p in hRECs in the LG and HG groups were measured by the quantitative real-time polymerase chain reaction. Transwell assay, Matrigel assay, and Scratch assay were used to analyze the migration, tube formation, and wound healing of hRECs. The immunofluorescence assay was used to analyze the effect of miR-615-5p on the expression of vascular endothelial growth factor A (VEGFA).Results　Compared with the LG group (1.00±0.28), the relative expression level of miR-615-5p in hRECs in the HG group (6.36±1.31) was higher (t=12.69, P<0.01). Transwell assay results showed that the relative numbers of cells migrated in the HG, mimic, mi-nc, inhibitor, and in-nc groups were 0.99±0.12, 0.96±0.08, 0.50±0.07, 0.72±0.13, and 0.96±0.10, respectively. Compared with the mi-nc group, the migration of hRECs in the mimic group was significantly stronger (t=6.10, P<0.05), while compared with the in-nc group, the migration of hRECs in the inhibitor group was significantly weaker (t=7.21, P<0.05). Matrigel assay results showed that the relative tube lengths of VEGFA in the HG, mimic, mi-nc, inhibitor, and in-nc groups were 1.00±0.15, 1.24±0.13, 0.81±0.13, 0.40±0.05, and 0.86±0.04, respectively. Compared with the mi-nc group, the tube formation of hRECs in the mimic group was significantly increased (t=5.52, P<0.01), while compared with the in-nc group, the tube formation of hRECs in the inhibitor group was significantly reduced (t=17.80, P<0.01). Scratch assay results showed that the wound healing rates of hRECs in the HG, mimic, mi-nc, inhibitor, and in-nc groups were 0.71±0.01,1.00±0.01,0.64±0.02,0.56±0.05, and 0.54±0.04, respectively. Compared with the mi-nc group, the wound healing rate of hRECs in the mimic group was significantly higher (t=25.30, P<0.01). The immunofluorescence assay results showed that the relative fluorescence intensities of hRECs in the HG, mimic, mi-nc, inhibitor, and in-nc groups were 1.00±0.09, 1.35±0.10, 1.02±0.04, 0.77±0.06, and 0.84±0.06, respectively. Compared with the mi-nc group, the expression level of VEGFA in hRECs in the mimic group was higher (t=3.78, P<0.05).Conclusion　High glucose promotes the expression of miR-615-5p in hRECs, thus increasing the migration, tube formation, and wound healing of hRECs. VEGFA may be the downstream target of miR-615-5p.