Abstract
Unveiling the mechanism of miR-122-5p in the mediation of forkhead box O3 (FOXO3) in regards to cochlear hair cell damage provides an effective solution for the treatment of ear hearing disorders. An oxidative stress model using a mouse cochlear hair cell line (HEI-OC1) was established via hydrogen peroxide (H2O2). Then HEI-OC1 cells were transfected with miR-122-5p mimic, miR-122-5p inhibitor, and lentiviral vector FOXO3-WT/MUT. Cell viability and apoptosis rate were determined by MTT assay and flow cytometry. Reactive oxygen species (ROS) were observed by confocal laser scanning microscopy. Bcl-2, Bax, capase-3 and c-caspase-9 levels were quantified by western blot analysis and quantitative reverse transcription polymerase chain reaction (RT-qPCR). Enzyme-linked immunosorbent assay (ELISA) was used to detect superoxide dismutase (SOD) and malondialdehyde (MDA) levels, and flow cytometry was performed to measure the mitochondrial membrane potential levels. In the HEI-OC1 oxidative stress model after transfection, the miR-122-5p level was decreased, whereas the FOXO3 level was increased, Moreover, the increased FOXO3 level diminished the cell viability, but promoted cell apoptosis. Apart from this, the Bcl-2 level was downregulated, while levels of Bax, c-caspase-3, c-caspase-9, ROS and MDA were upregulated. Meanwhile, the mitochondrial membrane potential level was also elevated. Overexpression of miR-122-5p was able to partially offset the effects of FOXO3 in the H2O2-treated HEI-OC1 cells. Collectively, miR-122-5p restrained the decrease in HEI-OC1 cell viability and apoptosis induced by treatment with H2O2.