miRNA-7a-2-3p Inhibits Neuronal Apoptosis in Oxygen-Glucose Deprivation (OGD) Model

Zi-Bin Zhang; Ya-Xin Tan; Qiong Zhao; Liu-Lin Xiong; Jia Liu; Fei-Fei Xu; Yang Xu; Larisa Bobrovskaya; Xin-Fu Zhou; Ting-Hua Wang

doi:10.3389/fnins.2019.00016

miRNA-7a-2-3p Inhibits Neuronal Apoptosis in Oxygen-Glucose Deprivation (OGD) Model

Front Neurosci. 2019 Jan 23:13:16. doi: 10.3389/fnins.2019.00016. eCollection 2019.

Authors

Zi-Bin Zhang^{1

2}, Ya-Xin Tan¹, Qiong Zhao³, Liu-Lin Xiong⁴, Jia Liu⁵, Fei-Fei Xu⁶, Yang Xu⁶, Larisa Bobrovskaya⁴, Xin-Fu Zhou⁴, Ting-Hua Wang¹

Affiliations

¹ Institute of Neuroscience, Kunming Medical University, Kunming, China.
² Department of Anesthesiology, Qilu Hospital of Shandong University, Jinan, China.
³ Department of Anesthesiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.
⁴ School of Pharmacy and Medical Sciences, Faculty of Health Sciences, University of South Australia, Adelaide, SA, Australia.
⁵ Department of Laboratory Animal Science, School of Basic Medical Sciences, Kunming Medical University, Kunming, China.
⁶ Institute of Neurological Diseases, Department of Anesthesiology and Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, China.

Abstract

Neuronal apoptosis is a major pathological hallmark of the neonatal hypoxic-ischemic brain damage (HIBD); however, the role of miR-7a-2-3p in the regulation of HIBD remains unknown. The purpose of this study was to explore the possible roles of miR-7a-2-3p in brain injury using a hypoxia-ischemia model in rats and oxygen-glucose deprivation (OGD) model in vitro. Firstly, we established the hypoxia-ischemia (HI) model and verified the model using Zea Longa scores and MRI in rats. Next, the changes of miR-7a-2-3p were screened in the ischemic cortex of neonatal rats by qRT-PCR at 12, 48, and 96 h after HIBD. We have found that the expression of miR-7a-2-3p in the HI rats decreased significantly, compared with the sham group (P < 0.01). Then, we established the OGD model in PC12 cells, SH-SY5Y cells and primary cortical neurons in vitro and qRT-PCR was used to confirm the changes of miR-7a-2-3p in these cells after the OGD. In order to determine the function of miR-7a-2-3p, PC12 cells, SH-SY5Y cells and rat primary cortical neurons were randomly divided into normal, OGD, mimic negative control (mimic-NC) and miR-7a-2-3p groups. Then, Tuj1+ (neuronal marker) staining, TUNEL assay (to detect apoptotic cells) and MTT assay (to investigate cell viability) were performed. We have found that the number of PC12 cells, SH-SY5Y cells and cortical neurons in the miR-7a-2-3p groups increased significantly (P < 0.01) in comparison to the OGD groups. The survival of cortical neurons in the miR-7a-2-3p group was improved markedly (P < 0.01), while the apoptosis of neurons in the miR-7a-2-3p group was significantly decreased (P < 0.01), compared with the normal group. Lastly, we investigated the target genes of miR-7a-2-3p by using the prediction databases (miRDB, TargetScan, miRWalk, and miRmap) and verified the target genes with qRT-PCR in the HI rats. Bioinformatics prediction showed that Vimentin (VIM), pleiomorphic adenoma gene 1(PLAG1), dual specificity phosphatase 10 (DUSP10), NAD(P)H dehydrogenase, quinone 1 (NQO1) and tumor necrosis factor receptor superfamily member 1B (TNFRSF1B) might be the targets of miR-7a-2-3p and the qRT-PCR confirmed that VIM increased in the HI rats (P < 0.01). In conclusion, miR-7a-2-3p plays a crucial role in the hypoxic-ischemic injury, and is associated with regulation of VIM.

Keywords: Vimentin (VIM); miR-7a-2-3p; neonatal hypoxic-ischemic brain damage (HIBD); neuronal apoptosis; oxygen-glucose deprivation (OGD).