miRNA-146a在视网膜色素上皮细胞老化及老年性黄斑变性中的功能研究

发布时间：2018-03-24 22:34

本文选题：miRNA-a　切入点：视网膜色素上皮细胞　出处：《眼科新进展》2017年02期

【摘要】：目的检测miRNA-146a(miR-146a)在视网膜色素上皮细胞(retinal pigment epithelial cells,RPE)衰老及老年性黄斑变性(age-related macular degeneration,AMD)中的表达水平,并探讨其通过抑制VEGF-A表达,参与调控AMD病变进程的机制。方法qRT-PCR检测年龄为2个月、8个月、12个月、18个月和24个月小鼠的RPE及75岁晚期"湿"性AMD患者眼球中miR-146以及VEGF-A的mRNA表达水平。Western Blotting检测VEGF-A蛋白表达水平。在ARPE-19细胞系中转染miR-146a,通过qRT-PCR及Western Blotting检测miR-146a对VEGF-A mRNA及蛋白水平的影响。结果在自然衰老的RPE中,miR-146的表达呈现随年龄增加逐渐上升的趋势,与2个月的小鼠相比,在18个月及24个月的小鼠中miR-146a水平上升约8倍及24倍。而VEGF-A的表达呈现下降趋势,与对照相比,24个月的小鼠RPE中的VEGF-A相对水平由2个月时的1.5倍降为0.8倍。但是在AMD患者中,病灶部位的miR-146a较病灶旁侧表达下降14.5倍,VEGF-A表达上升。在ARPE-19细胞中过表达miR-146a抑制了VEGF-A的mRNA及蛋白表达水平。结论本实验结果将miR-146a与RPE衰老及AMD的发生联系在一起,未来有可能成为RPE衰老及AMD早期诊断的新的分子标记物,为制定临床治疗策略提供参考。
[Abstract]:Objective to investigate the expression of miRNA-146a miR-146a in retinal pigment epithelial cells (RPE) and age-related macular degenerationin retinal pigment epithelial cells (RPE), and to explore the inhibition of VEGF-A expression. Methods qRT-PCR was used to detect the mRNA expression level of miR-146 and VEGF-A in the eyes of mice aged 2 months, 8 months, 12 months, 18 months and 24 months, and in the eyes of 75 years old patients with "wet" AMD. Western Blotting was used to detect the mRNA expression of miR-146 and VEGF-A in the eyes of mice aged 2 months, 8 months, 12 months, 18 months and 24 months. The effect of miR-146a on VEGF-A mRNA and protein levels was detected by qRT-PCR and Western Blotting. The results showed that the expression of miR-146 increased with age in naturally aging RPE. Compared with the mice of 2 months, the level of miR-146a increased about 8-fold and 24-fold in 18-month and 24-month-old mice, while the expression of VEGF-A decreased. Relative levels of VEGF-A in RPE of 24-month-old mice decreased from 1.5 times to 0.8 times at 2 months, but in patients with AMD, the relative level of VEGF-A decreased to 0.8 times as compared with control. The expression of VEGF-A was decreased by 14.5-fold in the miR-146a of the lesion. Overexpression of miR-146a in the ARPE-19 cells inhibited the expression of VEGF-A mRNA and protein. Conclusion the results of this study link miR-146a with RPE senescence and AMD. It may be a new molecular marker for the early diagnosis of RPE and AMD.
【作者单位】：抚顺市眼病医院;中国科学院上海生命科学研究院;
【分类号】：R774

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