miR-125b在小鼠白内障模型与人晶状体上皮细胞凋亡模型中的调控作用
李阳, 姜凌峰, 廉飞玥, 樊保良, 邵雨双, 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:
2019-12-20出版日期:
2020-09-30发布日期:
2020-09-15通讯作者:
秦宇E-mail:qinyubb@126.com作者简介:
李阳(1994-),男,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: 微小RNA-125b, BAK1, 凋亡, 白内障
Abstract: Objective To investigate the mechanism of targeting regulatory BAK1 protein by microRNA-125b (miR-125b) in an UV-induced mouse cataract model and a human lens epithelial cell apoptosis model. Methods LipofectamineTM RNAiMAX was used to transfect either miR-125b mimics or control RNAs,along with miR-125b inhibitor molecules and inhibitor controls to human lens epithelial cell line SRA01/04,either to up-regulate or down-regulate expression levels of miR-125b,respectively. After 48 h,ultraviolet UVB light (360 μW/cm2) was used to irradiate for 25 min to establish an apoptosis model. The cataract model was generated by irradiating the left eyes of mice with UVB light (360 μW/cm2),and the lens capsules of mice were collected. Transfection efficiencies were verified by real-time qPCR,and the expression levels of miR-125b and mRNA of its predictive target gene BAK1 were detected. The expression of BAK1 protein was detected by western blotting analysis,and the apoptosis rate of human lens epithelial cells was detected by TUNEL apoptosis assay. Results The results indicated that when compared with normal lens epithelial cells,the expression level of miR-125b in the apoptosis model of human lens epithelial cells was significantly decreased,along with observed increases in the expression of BAK1 protein and the apoptosis rate in human lens epithelial cells. Also,in the cells transfected with miR-125b mimics,the expression levels of miR-125b was significantly increased,the expression of BAK1 protein was decreased,and the apoptosis rate was decreased when compared to cells transfected with control RNA. In cells transfected with miR-125b inhibitor,the expression of miR-125b was significantly decreased,expression of BAK1 protein was significantly increased,and apoptosis was increased,when compared to the cells transfected with miR-125b inhibitor controls. Finally,a comparison between non-irradiated and UVB-irradiated mice models were made and the expression of miR-125b in the lens capsules was significantly decreased,expression of BAK1 protein was increased,and apoptosis was increased in the UVB-irradiated mice model group. Conclusion miR-125b affects the apoptosis of human lens epithelial cells through targeted regulation of BAK1,thus regulating the pathogenesis of cataract,and this could become a new strategy for non-operative targeted treatment of cataracts.
Key words: microRNA-125b, BAK1, apoptosis, cataract
PDF全文下载地址:
https://journal.cmu.edu.cn/CN/article/downloadArticleFile.do?attachType=PDF&id=2589