[1]陆博,马立威,王欣玲,等.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.[doi:10.13389/j.cnki.rao.2018.0024]
 LU Bo,MA Li-Wei,WANG Xin-Ling,et al.Expression of miR-138 and its effects on human lens epithelial cells apoptosis in age-related cataract patients[J].Recent Advances in Ophthalmology,2018,38(2):111-115.[doi:10.13389/j.cnki.rao.2018.0024]
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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/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
38卷
期数:
2018年2期
页码:
111-115
栏目:
实验研究
出版日期:
2018-02-05

文章信息/Info

Title:
Expression of miR-138 and its effects on human lens epithelial cells apoptosis in age-related cataract patients
作者:
陆博马立威王欣玲冯莉江陵峰王春霞张劲松阎启昌
110005 辽宁省沈阳市,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,中国医科大学眼科医院
Author(s):
LU BoMA Li-WeiWANG Xin-LingFENG LiJIANG Ling-FengWANG Chun-XiaZHANG Jin-SongYAN Qi-Chang
Department of Ophthalmology,the Fourth Affiliated Hospital of China Medical University,Key Labora-tory of Lens Research of Liaoning Province,Eye Hospital of China Medical University,Shenyang 110005,Liaoning Province,China
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
Keywords:
miR-138SIRT1age-related cataractapoptosisproliferation
分类号:
R776.1
DOI:
10.13389/j.cnki.rao.2018.0024
文献标志码:
A
摘要:
目的 检测miR-138在年龄相关性白内障晶状体组织中的表达情况,并探讨miR-138对人晶状体上皮细胞增殖和凋亡的影响及其可能的靶基因。方法 采用实时荧光定量PCR(real-time quantitative polymerase chain reaction,RT-qPCR)检测年龄相关性白内障患者(白内障组)与正常对照组中miR-138和预测靶基因沉默信息调节因子1(silent information regulator 1,SIRT1)的表达水平。向人晶状体上皮细胞系(SRA01/04)细胞中分别转染miR-138 模拟物、模拟物阴性对照物、miR-138抑制物、抑制物阴性对照物,采用RT-qPCR检测SIRT1的mRNA表达,Western blotting检测SIRT1的蛋白表达水平;转染72 h后细胞暴露于200 μmol·L-1 H2O2 1 h,Caspase-3活性检测试剂盒检测Caspase-3活性。双荧光素酶报告基因检测验证miR-138与SIRT1的靶向关系。结果 与正常对照组相比,白内障组miR-138的表达(3.64±0.19)显著升高(P<0.001),SIRT1的mRNA表达(0.32±0.06)显著下降(P<0.001);相对于模拟物阴性对照组,miR-138 模拟物组的SIRT1的mRNA(0.42±0.05)及蛋白(0.46±0.05)表达水平均明显降低,Caspase-3活性(3.24±0.17)明显升高(均为P<0.05);相对于抑制物阴性对照组,miR-138抑制物组的SIRT1的mRNA(2.95±0.13)及蛋白(1.98±0.12)表达水平均明显升高,Caspase-3活性(0.42±0.05)均明显下降(均为P<0.05);双荧光素酶报告基因检测确证SIRT1为miR-138的直接作用靶点。结论 miR-138在年龄相关性白内障患者晶状体囊膜中高表达,miR-138通过靶向性负性调控SIRT1表达,促进晶状体上皮细胞凋亡。
Abstract:
Objective To detect the expression of miR-138 in lens tissues of age-related 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 finally,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.

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备注/Memo

备注/Memo:
国家自然科学基金资助(编号:81170836、81570838);辽宁省自然科学基金资助(编号:2015020474)
更新日期/Last Update: 2018-02-02