MiR-212 Attenuates MPP+-Induced Neuronal Damage by Targeting KLF4 in SH-SY5Y Cells.
10.3349/ymj.2018.59.3.416
- Author:
Yanfeng SONG
1
;
Ying LIU
;
Xiaowei CHEN
Author Information
1. Department of Internal Medicine-Neurology, Hua Mei Branch of the Second People's Hospital of Liaocheng, Linqing, China. chenxiaoweicxww@163.com
- Publication Type:Original Article
- Keywords:
Parkinson's disease;
miR-212;
KLF4;
Notch signaling pathway;
SH-SY5Y cells
- MeSH:
1-Methyl-4-phenylpyridinium;
Apoptosis;
Blotting, Western;
Caspase 3;
Cell Count;
Cell Survival;
Enzyme-Linked Immunosorbent Assay;
Flow Cytometry;
In Vitro Techniques;
Interleukin-1beta;
L-Lactate Dehydrogenase;
Necrosis;
Nervous System Diseases;
Neurodegenerative Diseases;
Neurons*;
Parkinson Disease;
Reactive Oxygen Species;
RNA, Messenger;
Superoxide Dismutase
- From:Yonsei Medical Journal
2018;59(3):416-424
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: Parkinson's disease (PD) is a common age-dependent neurodegenerative disease. MiR-212 has been demonstrated to exert protective effects in several neurological disorders. The present study aimed to investigate the role and underlying molecular mechanism of miR-212 in PD. MATERIALS AND METHODS: 1-methyl-4-phenylpyridinium (MPP+)-induced SH-SY5Y cells were applied as a PD model in vitro. RTqPCR was used to measure the expression of miR-212 and Kruppel-like factor 4 (KLF4) mRNA. Western blot analysis was performed to detect the protein levels of KLF4, Notch1 and Jagged1. Cell viability and apoptosis were determined by the Cell Counting Kit-8 and flow cytometry, respectively. Quantitative analysis of caspase-3 activity, lactate dehydrogenase (LDH), reactive oxygen species (ROS), superoxide dismutase (SOD), tumor necrosis factor-α (TNF-α), and interleukin-1 beta (IL-1β) was conducted with corresponding ELISA kits. Dual-luciferase reporter assay was employed to evaluate the relationship between miR-212 and KLF4. RESULTS: MiR-212 was downregulated in MPP+-induced SH-SY5Y cells. Also, miR-212 alleviated MPP+-induced SH-SY5Y cell damage, embodied by increased cell viability, decreased caspase-3 activity, LDH release, ROS production, TNF-α, and IL-1β expression, as well as elevated SOD levels. KLF4 was a direct target of miR-212, and miR-212 repressed KLF4 expression in a post-transcriptional manner. Moreover, miR-212-mediated protection effects were abated following KLF4 expression restoration in MPP+-induced SH-SY5Y cells, represented as lowered cell viability and enhanced apoptotic rate. Furthermore, Notch signaling was involved in the regulation of miR-212/KLF4 axis in MPP+-induced SH-SY5Y cells. CONCLUSION: miR-212 might attenuate MPP+-induced neuronal damage by regulating KLF4/Notch signaling pathway in SH-SY5Y cells, a promising target for PD therapy.
