Effect of miR-129-5p on Neuronal Apoptosis and Oxidative Stress Induced by Oxygen Glucose Deprivation/Reoxygenation by Targeting SOX6
HUO Jiangtao, CAI Jinjin, LIANG Hui, GUO Gongbing, ZHANG Xiaoqiao*
The purpose of this study was to investigate the effect of microRNA-129-5p (miR-129-5p) targeting SOX6 (SRY-box transcription factor 6) on OGD/R (oxoglucose deprivation/reoxygenation)-induced apoptosis and oxidative stress in neuronal cells. Human neuronal cells (SH-SY5Y cells) were separated into control group, OGD/R group, miR-NC group, miR-129-5p mimics group, miR-129-5p mimics+pcDNA-NC group, and miR-129- 5p mimics+pcDNA-SOX6 group. Except for the control group, all other groups were induced with OGD/R. RT-qPCR was applied to detect the expression levels of miR-129-5p and SOX6 mRNA in cells. CCK-8 assay kit was used to detect cell proliferation rate. Flow cytometry was used to detect cell apoptosis. ELISA kit was used to detect the levels of intracellular SOD (superoxide dismutase), CAT (catalase), and GPx (glutathione peroxidase). H2DCFDA fluorescent probe was applied to detect cellular ROS (reactive oxygen species) level. Western blot was applied to detect the expression of SOX6, Caspase-3, and NF-κB proteins in cells. The results of the study are derived compared with the control group, the expression of miR-129-5p, cell activity, SOD, CAT, and GPx activities in OGD/R group decreased (P<0.05), while the expression of SOX6 mRNA, apoptosis rate, level of ROS, the expression of SOX6, Caspase-3, and p-NF-κB/NF-κB proteins increased (P<0.05). Compared with the OGD/R group, the expression of miR-129- 5p, cell activity, SOD, CAT, and GPx activities in miR-129-5p mimics group increased (P<0.05), while the expression of SOX6 mRNA, apoptosis rate, level of ROS, the expression of SOX6, Caspase-3, and p-NF-κB/NF-κB proteins decreased (P<0.05). Compared with the miR-129-5p mimics group, the expression of miR-129-5p, cell activity, SOD, CAT, and GPx activities in miR-129-5p mimics+pcDNA-SOX6 group decreased (P<0.05), while the expression of SOX6 mRNA, apoptosis rate, level of ROS, the expression of SOX6 and p-NF-κB/NF-κB proteins increased (P<0.05). Dual luciferase reporter gene experiment confirmed that SOX6 was the target gene of miR129-5p. Therefore, overexpression of miR-129-5p can downregulate SOX6 and alleviate OGD/R-induced neuronal apoptosis and oxidative stress.
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