Downregulated miRNA-324-5p aggravates neuronal injury induced by oxygen-glucose deprivation via modulating RAN

Gu,Junquan; Gui,Shuhua; Hu,Linlin; Kong,Liang; Di,Meiqi; Wang,Yinming

doi:10.3892/etm.2019.8249

Downregulated miRNA-324-5p aggravates neuronal injury induced by oxygen-glucose deprivation via modulating RAN

Authors:
- Junquan Gu
- Shuhua Gui
- Linlin Hu
- Liang Kong
- Meiqi Di
- Yinming Wang
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Affiliations: Department of Neurology, The Third People's Hospital of Wuxi, Wuxi, Jiangsu 214041, P.R. China
Published online on: November 27, 2019 https://doi.org/10.3892/etm.2019.8249
Pages: 658-664
Copyright: © Gu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Differentially expressed miRNAs in the GEO profile of ischemic stroke were analyzed to clarify the specific role of microRNA‑324‑5p (miRNA‑324‑5p) in ischemic stroke and the potential mechanism. After screening out miRNA‑324‑5p, its level in peripheral blood of stroke patients and in vitro oxygen‑glucose deprivation (OGD)‑induced primary rat neurons was determined by quantitative real‑time polymerase chain reaction (qRT‑PCR). Regulatory effects of miRNA‑324‑5p on viability, and apoptosis of OGD‑induced neurons were evaluated by CCK‑8 and Annexin V fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining, respectively. Glucose uptake and caspase-3 activity in OGD‑induced neurons transfected with miRNA‑324‑5p mimics or inhibitor were also examined. The binding of miRNA‑324‑5p to its target gene RAN was analyzed by dual‑luciferase reporter gene assay and western blot analysis. By analyzing the data of GSE46266 profile, miRNA‑324‑5p expression was shown markedly lower in MCAO rats relative to controls. Identically, we also observed the downregulated miRNA‑324‑5p in peripheral blood of stroke patients and in vitro OGD‑induced primary neurons. Overexpression of miRNA‑324‑5p accelerated viability, induced apoptosis and strengthened glucose uptake ability of OGD‑induced neurons. Knockdown of miRNA‑324‑5p, conversely, obtained the opposite results. Furthermore, we confirmed the binding of miRNA‑324‑5p to RAN, the target gene that was negatively regulated by miRNA‑324‑5p. Importantly, RAN overexpression partially reversed the regulatory effect of miRNA‑324‑5p on viability and glucose uptake of OGD‑induced neurons. miRNA‑324‑5p is downregulated after ischemic stroke, which aggravates the disease condition by inhibiting neuronal proliferation and glucose uptake via upregulating RAN.

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