MicroRNA-486 targeting TRIM10 to inhibit Parkinson’s disease cell model damage

doi:10.16098/j.issn.0529-1356.2022.04.004

Abstract

Abstract:

Objective To study the effect and mechanism of microRNA-486(miR-486) on 1-methyl-4-phenylpyridine (MPP+)-induced apoptosis of Parkinson’s disease (PD) PC12 cells in vitro. Methods The experiment was divided into control group, PD (MPP+ induced PC12 cells), miR-NC (transfected mimics control, MPP+ induced PC12 cells), miR-486 group (transfected miR-486 mimics, MPP+ induced PC12 cells), miR-486+vector group (co-transfected miR-486 mimics, pcDNA3.1), miR-486+TRIM10 group (co-transfected miR-486 mimics, pcDNA3.1-TRIM10)，n=9 each group. CCK-8 method was used to analyze cell proliferation changes, flow cytometry was used to analyze cell apoptosis levels, Western blotting was used to analyze changes in Bax and Bcl-2 protein expression, thiobarbituric acid method was used to detect malondialdehyde (MDA) content in cells, fluorescence probe method was used to detect the level of reactive oxygen species (ROS) in the cells, and 2, 4-dinitrophenylhydrazine was used to analyze the level of lactate dehydrogenase (LDH) in the culture supernatant. Bioinformatics software was used to predict the target genes of miR-486, and the detection of targeting relationship between imiR-486 and TRIM 10 by dual luciferase reporter gene assay. Results Compared with the control group, the cell survival rate and Bcl-2 protein expression in the PD group decreased, while the apoptosis rate, Bax protein expression, MDA, ROS and LDH levels increased. Compared with the miR-NC group, the cell survival rate and Bcl-2 protein expression in the miR-486 group were increased, and the apoptosis rate, Bax protein expression, MDA, ROS and LDH levels were decreased. MiR-486 targeted down-regulation of TRIM10 expression. Compared with the miR-486+vector group, the miR-486+TRIM10 group decreased the cell survival rate and Bcl-2 protein level, while the apoptosis rate, Bax protein level, MDA, ROS and LDH levels increased. Conclusion Up-regulation of miR-486 targeted and inhibited TRIM10 to reduce MPP+induced apoptosis in in vitro Parkinson’s PC12 cell models.

Key words: MicroRNA-486, TRIM10, Parkinson’s disease, Western blotting, PC12 cell

CLC Number:

R741.02

LIAO Jian-yun JIANG Xin-liu XIE Wei-fa. MicroRNA-486 targeting TRIM10 to inhibit Parkinson’s disease cell model damage[J]. Acta Anatomica Sinica, 2022, 53(4): 424-431.

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