microRNA-125b通过调控Nrf2/Keap1信号通路影响光感受器细胞氧化应激
刘金霞, 王钰池, 郭卓, 赵江月, 孔珺, 秦宇中国医科大学附属第四医院眼科, 中国医科大学眼科医院, 辽宁省晶状体学重点实验室, 沈阳 110005
收稿日期:
2021-01-27出版日期:
2021-11-30发布日期:
2021-11-04通讯作者:
秦宇E-mail:qinyubb@126.com作者简介:
刘金霞(1995-),女,硕士研究生.基金资助:
国家自然科学基金青年基金(81600717);辽宁省自然科学基金(201602851)关键词: microRNA-125b, 核因子E2相关因子, 氧化应激, 干性年龄相关性黄斑变性
Abstract: Objective To investigate the regulating mechanism of microRNA-125b (miR-125b) in photoreceptor cells via the Nrf2/Keap1 signaling pathway using an H2O2-induced oxidative stress model. Methods The oxidative stress model was established using the mouse cone cell line 661W treated with H2O2 (800 μmol/L) for 6 h. Expression of miR-125b and Keap1/Nrf2/HO-1 was detected using real-time quantitative PCR (qPCR). miR-125b mimic and mimic control and miR-125b inhibitor and inhibitor control were transfected into the treated 661W cells in serum-free Opti-MEM using LipofectamineTM RNAiMAX. miR-125b expression, 48 hours post-transfection, in 661W cells was up- and downegulated, respectively. Expression of Keap1 protein was detected using Western blotting. Results Compared to the control group, expression of miR-125b decreased significantly, that of Nrf2 and its downstream gene HO-1 increased significantly, and that of Keap1 decreased significantly in the H2O2-treated group. Furthermore, after transfection with miR-125b mimic, the mRNA expression of Nrf2, HO-1, and miR-125b increased significantly. However, Keap1 mRNA expression showed no significant change, whereas Keap1 protein expression decreased significantly. Conversely, after transfection with miR-125b inhibitor, mRNA expression of Nrf2, HO-1, and miR-125b was significantly downregulated and that of Keap1 showed no significant change, whereas Keap1 protein expression level significantly increased. All differences were significant (P < 0.05). Conclusion The miR-125b/Keap1/Nrf2/HO-1 axis is a potential regulatory mechanism in pathogenesis and treatment of dry age-related macular degeneration that witnesses severe oxidative stress and adversely damaged photoreceptor cells.
Key words: microRNA-125b, nuclear factor-erythroid2-related factor 2, oxidative stress, dry age-related macular degeneration
PDF全文下载地址:
https://journal.cmu.edu.cn/CN/article/downloadArticleFile.do?attachType=PDF&id=2871