眼科新进展2018，Vol.38Issue(2)：111-115,5.DOI:10.13389/j.cnki.rao.2018.0024
年龄相关性白内障患者miR-138的表达及其对人晶状体上皮细胞凋亡的影响
Expression of miR-138 and its effects on human lens epithelial cells apoptosis in age-related cataract patients
摘要
Abstract
Objective To detect the expression of miR-138 in lens tissues of agerelated cataract and explore the effects of miR-138 on the proliferation and apoptosis of human lens epithelial cells and its possible target genes.Methods Real-time quantitative PCR (RT-qPCR) was applied for the detection of the expression of miR-138 and prediction of target gene sirtuin (silent information regulator 1) (SIRT1) in patients with age-related cataract (cataract group) and anterior lens capsules (normal control group).Then miR-138 mimics,mimic controls,miR-138 inhibitors and inhibitor controls were transfected into the human lens epithelial cell line (SRA01/04),and the expression of SIRT1 mRNA and protein was detected by RT-qPCR and Western blot,accordingly.At 72 hours after transfection,the cells were exposed to 200 μmol · L-1 H2O2 for 1 hour,followed by detection of the activity of Caspase-3 by the Caspase-3 activity assay kit,and identification of the targeted relationship between miR-138 and SIRT1 by dual luciferase reporter assays.Results Compared with the normal control group,the expression of miR-138(3.64 ±0.19) was significantly increased (P ＜0.001),but the expression of SIRT1 mRNA(0.32 ± 0.06) was significantly decreased (P ＜ 0.001) in the cataract group.Moreover,The expression levels of SIRT1 mRNA(0.42 ± 0.05) and protein(0.46 ± 0.05) in cells transfected with miR-138 mimics were significantly decreased,while the activity of Caspase-3 (3.24 ± 0.17) was significantly elevated when compared with cells transfected with minic controls (all P ＜ 0.05);Compared with cells transfected with inhibitor controls,the expressions of SIRT1 mRNA(2.95 ±0.13) and protein(1.98 ±0.12) were significantly upregulated,whereas Caspase-3 activity(0.42 ±0.05) was significantly decreased in cells transfected with miR-138 inhibitors (all P ＜0.05).And fmally,dual luciferase reporter assays showed the confirmation SIRT1 as a direct target of miR-138.Conclusion miR-138 is highly expressed in the lens capsule of age-related cataract patients,and it can promote the apoptosis of lens epithelial cells by negatively regulating the expression of SIRT1.关键词
miR-138/沉默信息调节因子1/年龄相关性白内障/凋亡/增殖Key words
miR-138/SIRT1/age-related cataract/apoptosis/proliferation分类
医药卫生引用本文复制引用
陆博,马立威,王欣玲,冯莉,江陵峰,王春霞,张劲松,阎启昌..年龄相关性白内障患者miR-138的表达及其对人晶状体上皮细胞凋亡的影响[J].眼科新进展,2018,38(2):111-115,5.基金项目
国家自然科学基金资助(编号:81170836、81570838) （编号:81170836、81570838）
辽宁省自然科学基金资助(编号:2015020474)National Natural Science Foundation of China (No:81170836,81570838) （编号:2015020474）
Natural Science Foundation of Liaoning Province (No:2015020474) （No:2015020474）
