miR-138调控年龄相关性白内障晶状体上皮细胞抗氧化应激作用的机制  

Effect of miR-138 on the antioxidant function of lens epithelial cells affected by age-related cataracts

作　　者：陆博[1] 马立威[1] 王欣玲[1] 韩笑[1] 冯莉[1] 姜凌峰[1] 王春霞[1] 张劲松[1] 阎启昌[1] Bo Lu;Li-Wei Ma;Xin-Ling Wang;Xiao Han;Li Feng;Ling-Feng Jiang;Chun-Xia Wang;Jin-Song Zhang;Qi-Chang Yan(Department of Ophthalmology,the Fourth Affiliated Hospital of China Medical University;Key Laboratory of Lens Research of Liaoning Province;Eye Hospital of China Medical University,Shenyang 110005,Liaoning Province,China)

机构地区：[1]中国医科大学附属第四医院眼科,辽宁省晶状体学重点实验室,中国医科大学附属眼科医院,中国辽宁省沈阳市110005

出　　处：《国际眼科杂志》2018年第4期610-614,共5页International Eye Science

基　　金：国家自然科学基金资助项目(No.81170836,81570838)

摘　　要：目的:探讨miR-138对年龄相关性白内障晶状体上皮细胞抗氧化应激能力的影响及其作用机制。方法:采用实时定量PCR(RT-q PCR)检测年龄相关性白内障与正常人透明晶状体前囊膜组织中及人晶状体上皮细胞系(SRA01/04)氧化应激模型中miR-138的表达水平。利用2',7'-二氯荧光黄双乙酸盐(DCFH-DA)荧光探针检测细胞内源性活性氧(reactive oxygen species,ROS)水平。向人晶状体上皮细胞中分别转染miR-138 mimics,mimic controls,miR-138 inhibitors,inhibitor controls 72h后细胞暴露于400μmol/L H_2O_2 1h,采用RT-qPCR检测p53和Bax的mRNA表达,western blotting检测p53和Bax的蛋白表达水平,MTS法检测细胞增殖活力。结果:与正常对照组相比,年龄相关性白内障晶状体组织中与人晶状体上皮细胞氧化应激模型中miR-138的表达均显著升高,差异有统计学意义(P<0.001);人晶状体上皮细胞氧化应激模型中内源性ROS的水平明显升高,差异有统计学意义(P<0.001)。相对于对照组,miR-138 mimics组p53和Bax的mRNA、蛋白表达水平均明显升高,细胞增殖活力明显下降,差异有统计学意义(均P<0.001);miR-138 inhibitors组p53和Bax的mRNA、蛋白表达水平均明显下降,细胞增殖活力显著升高,差异有统计学意义(均P<0.001)。结论:miR-138在年龄相关性白内障囊膜组织中表达上调,通过正调控下游靶基因p53和Bax,下调人晶状体上皮细胞抗氧化应激的能力,抑制人晶状体上皮细胞增殖和修复,从而参与年龄相关性白内障的发生过程。AIM:To investigate the effects and mechanism of miR-138 in mediating the antioxidant function of lens epithelial cells affected by age-related cataracts.METHODS:Real-time quantitative PCR(RT-qPCR)was used to detect miR-138 expression in the anterior lens capsules of healthy people,the anterior lens capsules of patients with age-related cataracts,and human epithelial cell line(SRA01/04)cells exposed to oxidative stress.A 2',7'-dichloro-fluorescein diacetate(DCFH-DA)probe was used to measure the levels of endogenous reactive oxygen species(ROS)in human lens epithelial cells(hLECs)exposed to 400mol/L H2 O2 for 1h.SRA01/04 cells were transfected with either miR-138 mimics,mimic controls,miR-138 inhibitors or inhibitor controls.After 72h,these cells were exposed to 400mol/L H2 O2 for 1h,then p53 and Bax mRNA expression were measured using RT-qPCR.Expression of p53 and Bax protein were also measured by western blotting analysis.Finally,cell viability was assessed using an MTS assay.RESULTS:Compared to the control group,expression of miR-138 in the anterior lens capsules of age-related cataract patients and in SRA01/04 cells exposed to oxidative stress significantly increased(P<0.001).Levels of endogenous ROS were significantly elevated in hLECs exposed to oxidative stress(P<0.001).Compared to the mimic control group,the hLECs in the miR-138 mimic group expressed significantly higher levels of p53 and Bax mRNA and protein while cell viability was significantly reduced(P<0.001).Conversely,p53 and Bax mRNA and protein expression were significantly reduced in the miR-138 inhibitor group as compared to the control group,while the cells in this group had much higher levels of cell viability(P<0.001).CONCLUSION:The expression of miR-138 is upregulated in the anterior lens capsules of age-related cataract patients.MiR-138 decreases the anti-oxidative stress capacity of lens epithelial cells by upregulating p53 and Bax,while inhibiting cell proliferation and repair.This finding suggests that miR-138 may play a key role in the d

关 键 词：miR-138 P53 BAX 年龄相关性白内障 增殖 

分 类 号：R776.1[医药卫生—眼科]