CircDLGAP4 overexpression relieves oxygen-glucose deprivation-induced neuronal injury by elevating NEGR1 through sponging miR-503-3p

Circular RNAs (circRNAs) have been reported to play vital regulatory roles in human diseases. However, the functions of circRNAs in ischemic stroke (IS) are limited. In this study, we aimed to explore the functions and mechanisms of circRNA DLG associated protein 4 (circDLGAP4) in IS development. Ox...

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Published in:	Journal of molecular histology 2022-04, Vol.53 (2), p.321-332
Main Authors:	Qiu, Lingling, He, Jinfeng, Chen, Hui, Xu, Xiaohui, Tao, Yongjun
Format:	Article
Language:	eng
Subjects:	Apoptosis Biomedical and Life Sciences Biomedicine Cell Adhesion Molecules, Neuronal - metabolism Cell Biology Cell culture Cell cycle Cell death Cell viability Cholecystokinin Chromosome 3 Cytokines Developmental Biology Enzyme-linked immunosorbent assay Flow cytometry Gene expression Glucose Glucose - metabolism GPI-Linked Proteins - metabolism Growth regulators Histology Humans Immunoprecipitation Inflammation Ischemia Life Sciences MicroRNAs MicroRNAs - genetics MicroRNAs - metabolism miRNA mRNA Neurons - metabolism Original Paper Oxygen Oxygen - metabolism Polymerase chain reaction Proteins Ribonuclease RNA, Circular - genetics SAP90-PSD95 Associated Proteins Stroke
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Summary:	Circular RNAs (circRNAs) have been reported to play vital regulatory roles in human diseases. However, the functions of circRNAs in ischemic stroke (IS) are limited. In this study, we aimed to explore the functions and mechanisms of circRNA DLG associated protein 4 (circDLGAP4) in IS development. Oxygen–glucose deprivation (OGD)-treated HCN-2 cells were used to mimic IS environment in vitro. Quantitative real-time polymerase chain reaction (qRT-PCR) assay was used to detect the levels of circDLGAP4, microRNA-503-3p (miR-503-3p) and neuronal growth regulator 1 (NEGR1) mRNA. RNase R assay was conducted to analyze the stability of circDLGAP4. Cell Counting Kit-8 (CCK-8) assay and flow cytometry analysis were adopted for cell viability and death, respectively. Western blot assay was performed for protein levels. Enzyme-linked immunosorbent assay (ELISA) kits were used to examine the concentrations of inflammatory cytokines. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull-down assay were employed to analyze the relationships among circDLGAP4, miR-503-3p and NEGR1. CircDLGAP4 level was declined in HCN-2 cells after OGD treatment. CircDLGAP4 overexpression promoted cell viability and suppressed cell death and inflammatory cytokine concentrations in OGD-treated HCN-2 cells. CircDLGAP4 acted as the sponge for miR-503-3p and the impacts of circDLGAP4 overexpression on cell viability, death and inflammation in OGD-treated HCN-2 cells were reversed by miR-503-3p elevation. Furthermore, NEGR1 was the target gene of miR-503-3p. MiR-503-3p inhibition ameliorated OGD-induced HCN-2 cell impairments, but NEGR1 knockdown abolished the effects. CircDLGAP4 alleviated OGD-induced HCN-2 cell damage by regulating miR-503-3p/NEGR1 axis.
ISSN:	1567-2379 1567-2387