Upregulation of miR-376c-3p alleviates oxygen\u2013glucose deprivation-induced cell injury by targeting ING5

Heng Zhang,Jie Zhou,Mingxia Zhang,Bing He,Yanjie Yi

doi:10.1186/s11658-019-0189-2

Heng Zhang, Jie Zhou + Show 3 more

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https://doi.org/10.1186/s11658-019-0189-2

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Abstract

BackgroundThe expression level of miR-376c-3p is significantly lower in infants with neonatal hypoxic-ischemic encephalopathy (HIE) than in healthy infants. However, the biological function of this microRNA remains largely elusive.MethodsWe used PC-12 and SH-SY5Y cells to establish an oxygen–glucose deprivation (OGD) cell injury model to mimic HIE in vitro. The miR-376c-3p expression levels were measured using quantitative reverse transcription PCR. The CCK-8 assay and flow cytometry were utilized to evaluate OGD-induced cell injury. The association between miR-376c-3p and inhibitor of growth 5 (ING5) was validated using the luciferase reporter assay. Western blotting was conducted to determine the protein expression of CDK4, cyclin D1, Bcl-2 and Bax.ResultsMiR-376c-3p was significantly downregulated in the OGD-induced cell injury model. Its overexpression elevated cell viability and impaired cell cycle G0/G1 phase arrest and apoptosis in PC-12 and SH-SY5Y cells after OGD. Downregulation of miR-376c-3p gave the opposite results. We further demonstrated that ING5 was a negatively regulated target gene of miR-376c-3p. Importantly, ING5 knockdown had a similar effect to miR-376c-3p-mediated protective effects against cell injury induced by OGD. Its overexpression abolished these protective effects.ConclusionOur data suggest that miR-376c-3p downregulated ING5 to exert protective effects against OGD-induced cell injury in PC-12 and SH-SY5Y cells. This might represent a novel therapeutic approach for neonatal HIE treatment.

Highlights

The expression level of miR-376c-3p is significantly lower in infants with neonatal hypoxic-ischemic encephalopathy (HIE) than in healthy infants
The levels of miR-376c-3p decrease in the oxygen–glucose deprivation (OGD)-induced cell injury model PC-12 and SH-SY5Y cells in an OGD model were used to investigate the potential role of miR-376c-3p in HIE brain injury
The Cell Counting Kit-8 (CCK-8) assay showed that the cell viability of PC-12 and SH-SY5Y cells decreased significantly after OGD (Fig. 1b, p < 0.01)

Summary

Introduction

The expression level of miR-376c-3p is significantly lower in infants with neonatal hypoxic-ischemic encephalopathy (HIE) than in healthy infants. Neonatal hypoxic-ischemic encephalopathy (HIE), which is known as neonatal stroke, is caused by a disruption of cerebral blood vessels and leads to hypoxic or ischemic injury [1]. It is considered a major cause of disability in children after the neonatal period, with 0.1–0.2% incidence in term or near-term infants [2, 3]. Up to 40% of HIE patients usually suffer from devastating disability, including cerebral palsy, mental retardation, epilepsy and learning impairment [4,5,6]. It is important to better understand the molecular mechanisms of HIE.

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