MicroRNA-124 Prevents H2O2 Induced Apoptosis and Oxidative Stress in Human Lens Epithelial Cells via Inhibition of the NF-κB Signaling Pathway

被引:24
|
作者
Gu, Xiu-li [1 ]
机构
[1] Yantaishan Hosp, Dept Ophthalmol, 91 Jiefang Rd, Yantai 264000, Shandong, Peoples R China
关键词
MicroRNA-124; NF-kappa B; Human lens epithelial cells; Apoptosis; Oxidative stress; NUCLEAR-FACTOR; IN-VITRO; EXPRESSION; MIR-124; CATARACT; AGE; ACTIVATION; GENE; PATHOGENESIS; DYSFUNCTION;
D O I
10.1159/000491433
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Aim: To investigate the regulation of microRNA-124 (miRNA-124) on NF-kappa B pathway from H2O2-induced apoptosis and oxidative stress in human lens epithelial cells (hLEC). Methods: The MTT (3-[4, 5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide) assay was used to detect hLEC viability. HLECs were divided into Blank, H2O2, mimics (miRNA-124 mimics) + H2O2, NC+ H2O2, pyrrolidine dithiocarbamate (PDTC; NF-kappa B signaling pathway inhibitor) + H2O2, and inhibitors (miRNA-124 inhibitors) + PDTC + H2O2 groups. Quantitative real-time polymerase chain reaction and Western blot were employed to detect mRNA and protein expressions, Dichloro-dihydro-fluorescein diacetate to measure reactive oxygen species (ROS) production, and AnnexinV-FITC/PI staining to determine cell apoptosis. The mitochondrial membrane potential (MMP) was detected by fluorescence probe JC-1. Results: The H2O2-induced hLEC showed reductions in cell viability with decreased miRNA-124 but increased p-p65 in a dose-/time-dependent manner. Furthermore, ROS production, malondialdehyde content, Bax and Caspase-3 expressions, and cell apoptosis were elevated in H2O2-induced hLEC, whereas the activities of superoxide dismutase and glutathione peroxidase, Bcl-2 expression, MMP, as well as the mitochondrial energy metabolism genes were reduced. Additionally, miRNA-1 24 mimics and PDTC both decreased the p-p65 and reversed the cytotoxicity in H2O2-induced hLEC. Conclusion: MiRNA-124 prevents H2O2-induced oxidative stress and apoptosis in hLEC through suppressing the activation of the NF-kappa B pathway. (C) 2018 S. Karger AG, Basel
引用
收藏
页码:213 / 222
页数:10
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