Inhibition of miR-128 Abates Aβ-Mediated Cytotoxicity by Targeting PPAR-γ via NF-κB Inactivation in Primary Mouse Cortical Neurons and Neuro2a Cells.
10.3349/ymj.2018.59.9.1096
- Author:
Lijiao GENG
1
;
Tao ZHANG
;
Wei LIU
;
Yong CHEN
Author Information
1. Department of Rehabilitation Medicine, Huaihe Hospital of Henan University, Kaifeng, China. angelcindtg@yahoo.com
- Publication Type:Original Article
- Keywords:
Alzheimer's disease;
microRNA-128;
PPAR-γ;
NF-κB
- MeSH:
Alzheimer Disease;
Animals;
Blotting, Western;
Caspase 3;
Cause of Death;
Cell Survival;
Computational Biology;
Down-Regulation;
Flow Cytometry;
Humans;
Luciferases;
Mice*;
MicroRNAs;
Neurodegenerative Diseases;
Neurons*;
Plasma;
PPAR gamma;
RNA, Messenger;
Transcription Factor RelA;
United States;
Up-Regulation
- From:Yonsei Medical Journal
2018;59(9):1096-1106
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: Alzheimer's disease (AD) is the sixth most common cause of death in the United States. MicroRNAs have been identified as vital players in neurodegenerative diseases, including AD. microRNA-128 (miR-128) has been shown to be dysregulated in AD. This study aimed to explore the roles and molecular mechanisms of miR-128 in AD progression. MATERIALS AND METHODS: Expression patterns of miR-128 and peroxisome proliferator-activated receptor gamma (PPAR-γ) messenger RNA in clinical samples and cells were measured using RT-qPCR assay. PPAR-γ protein levels were determined by Western blot assay. Cell viability was determined by MTT assay. Cell apoptotic rate was detected by flow cytometry via double-staining of Annexin V-FITC/PI. Caspase 3 and NF-κB activity was determined by a Caspase 3 Activity Assay Kit or NF-κB p65 Transcription Factor Assay Kit, respectively. Bioinformatics prediction and luciferase reporter assay were used to investigate interactions between miR-128 and PPAR-γ 3′UTR. RESULTS: MiR-128 expression was upregulated and PPAR-γ expression was downregulated in plasma from AD patients and amyloid-β (Aβ)-treated primary mouse cortical neurons (MCN) and Neuro2a (N2a) cells. Inhibition of miR-128 decreased Aβ-mediated cytotoxicity through inactivation of NF-κB in MCN and N2a cells. Moreover, PPAR-γ was a target of miR-128. PPAR-γ upregulation attenuated Aβ-mediated cytotoxicity by inactivating NF-κB in MCN and N2a cells. Furthermore, PPAR-γ downregulation was able to abolish the effect of anti-miR-128 on cytotoxicity and NF-κB activity in MCN and N2a cells. CONCLUSION: MiR-128 inhibitor decreased Aβ-mediated cytotoxicity by upregulating PPAR-γ via inactivation of NF-κB in MCN and N2a cells, providing a new potential target in AD treatment.
