miR-873-5p micro-RNA protects neurons damaged by epilepsy by regulating voltage-dependent anion channel protein 1
10.3760/cma.j.issn.0254-1424.2021.02.002
- VernacularTitle:miR-873-5p靶向调控VDAC1保护大鼠癫痫样神经细胞的作用机制探讨
- Author:
Yongqiang GAO
;
Yan DONG
;
Xin CHENG
;
Minggao CHENG
;
Wenjing XU
;
Yongbin LANG
;
Jun WANG
- From:
Chinese Journal of Physical Medicine and Rehabilitation
2021;43(2):109-115
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the regulatory effect of miR-873-5p micro-RNA targeting voltage-dependent anion channel protein 1 (VDAC1) in neurons and its mechanism.Methods:Murine nerve cells were randomly divided in vitro into a control group, a model group, a mimetic negative carrier (miR-con) group and an miR-873-5p group. The epileptiform hippocampal nerve cell model was induced in all of the cells except those in the control group using magnesium-free medium. The control group was normally cultured, while the miR-con and miR-873-5p groups were transfected with miR control and miR-873-5p RNA respectively. Real-time fluorescent quantitative polymerase chain reactions were used to detect the expression of miR-873-5p and VDAC1 mRNA. Western blotting was employed to detect VDAC1, B-cell lymphoma/leukemia-2 protein (Bcl-2), Bcl-2 associated X protein (Bax) and cleared caspase-3 in the neurons. The levels of reactive oxygen species (ROS), malondialdehyde (MDA) and glutathione (GSH) were measured using the DCFH-DA fluorescent probe, the thiobarbituric acid method and enzyme-linked immunosorbent assay respectively. Any apoptosis was detected using flow cytometry, while the targeting of miR-873-5p on VDAC1 was verified using the double fluorescence zymase reporter gene method.Results:Compared with the control group, a significant decrease in the average expression of miR-873-5p, Bcl-2 and in GSH and MDA levels was observed in the model group, but there was a significant increase in the average level of VDAC1, Bax, cleaved caspase-3 and ROS and in the rate of apoptosis. Compared with the miR-con group, a significant decrease in the average expression of Bax, cleaved caspase-3, ROS and in the apoptosis rate was observed in the miR-873-5p group, but there was a significant increase in the average level of Bcl-2, GSH and MDA. Moreover, it was verified that miR-873-5p reduced the expression of VDAC1.Conclusion:miR-873-5p protects damaged neurons by inhibiting their apoptosis through negatively regulating the target gene VDAC1 and the oxidative stress response.
