Eaf2基因调控microRNA抑制白内障发生的机制研究

摘要

Objective To explore the expression of microRNA in Eaf2 knockout mice and the effect of Eaf2 on the apoptosis of human lens epithe⁃lial cells and the expression of microRNA in lens. Methods pEGFP⁃C1⁃Eaf2 was transfected into SRA01/04 cells using Lipofectamine 2000 to over express Eaf2 gene,and then the flow cytometry was used to detect cell apoptosis rate. And real time q⁃PCR was used to measure the expression of microRNA both in human lens epithelial cells and Eaf2 knockout mice. Results Compared with controls,the apoptosis rate of cells transfected with pEGFP⁃C1⁃Eaf2 was reduced,the expression of miR⁃125b and let⁃7a was significantly increased and miR⁃204 was decreased in cells transfect⁃ed with pEGFP⁃C1⁃Eaf2. Compared with controls,the expression of miR⁃125b and let⁃7a was lower and miR⁃204 was higher in Eaf2 knockout mice. Each result was statistically significant(P<0.01). Conclusion Eaf2 might inhibit apoptosis of human lens epithelial cells via regulating the expression of microRNA. Eaf2 may have a protective effect for the lens in the genesis of cataract.%目的：探讨Eaf2基因敲除小鼠中microRNA的表达情况，以及Eaf2基因对人晶状体上皮细胞凋亡及晶状体内microRNA表达的影响。方法利用Lipofectamine 2000瞬时转染pEGFP⁃C1⁃Eaf2质粒过表达Eaf2基因，利用流式细胞仪检测细胞凋亡率的变化，利用Real time q⁃PCR方法检测人晶状体上皮细胞中microRNA的表达情况；利用Real time q⁃PCR方法检测Eaf2基因敲除小鼠microRNA的表达情况。结果 pEGFP⁃C1⁃Eaf2质粒转染组细胞凋亡率显著低于对照组，miR⁃125b、let⁃7a的表达显著高于对照组，而miR⁃204的表达显著低于对照组；Eaf2基因敲除小鼠miR⁃125b、let⁃7a的表达显著低于对照组，而miR⁃204的表达显著高于对照组，差异均有统计学意义（P<0.01）。结论 Eaf2基因可能通过调控microRNA的表达抑制人晶状体上皮细胞凋亡，从而在白内障发生中发挥晶状体保护作用。