本文選題：miRNA-a　切入點(diǎn)：視網(wǎng)膜色素上皮細(xì)胞　出處：《眼科新進(jìn)展》2017年02期

【摘要】：目的檢測miRNA-146a(miR-146a)在視網(wǎng)膜色素上皮細(xì)胞(retinal pigment epithelial cells,RPE)衰老及老年性黃斑變性(age-related macular degeneration,AMD)中的表達(dá)水平,并探討其通過抑制VEGF-A表達(dá),參與調(diào)控AMD病變進(jìn)程的機(jī)制。方法qRT-PCR檢測年齡為2個月、8個月、12個月、18個月和24個月小鼠的RPE及75歲晚期"濕"性AMD患者眼球中miR-146以及VEGF-A的mRNA表達(dá)水平。Western Blotting檢測VEGF-A蛋白表達(dá)水平。在ARPE-19細(xì)胞系中轉(zhuǎn)染miR-146a,通過qRT-PCR及Western Blotting檢測miR-146a對VEGF-A mRNA及蛋白水平的影響。結(jié)果在自然衰老的RPE中,miR-146的表達(dá)呈現(xiàn)隨年齡增加逐漸上升的趨勢,與2個月的小鼠相比,在18個月及24個月的小鼠中miR-146a水平上升約8倍及24倍。而VEGF-A的表達(dá)呈現(xiàn)下降趨勢,與對照相比,24個月的小鼠RPE中的VEGF-A相對水平由2個月時的1.5倍降為0.8倍。但是在AMD患者中,病灶部位的miR-146a較病灶旁側(cè)表達(dá)下降14.5倍,VEGF-A表達(dá)上升。在ARPE-19細(xì)胞中過表達(dá)miR-146a抑制了VEGF-A的mRNA及蛋白表達(dá)水平。結(jié)論本實(shí)驗結(jié)果將miR-146a與RPE衰老及AMD的發(fā)生聯(lián)系在一起,未來有可能成為RPE衰老及AMD早期診斷的新的分子標(biāo)記物,為制定臨床治療策略提供參考。
[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.
【作者單位】： 撫順市眼病醫(yī)院;中國科學(xué)院上海生命科學(xué)研究院;
【分類號】：R774

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