MiR-130a exerts neuroprotective effects against ischemic stroke through PTEN/PI3K/AKT pathway

Tingting Zheng,Yu Shi,Jun Zhang,Jiao Peng,Xue Zhang,Keke Chen,Yun Chen,Li Liu

doi:10.1016/j.biopha.2019.109117

Abstract

BackgroundIschemic stroke is significantly affected by the dysfunction of the miRNA network. Recent research has described that disordered expression of miR-130a is associated with ischemic stroke. Here, we aimed to investigate the possible mechanism of the miR-130a-mediated neuroprotection that follows ischemia-reperfusion (I/R) injury. MethodThis study was comprised of two models: oxygen-glucose deprivation/Reperfusion (OGDR) and middle cerebral artery occlusion (MCAO). RT-PCR and immunoblotting were used to examine gene expression levels, and MTT assay and flow cytometric analysis were used to examine cell states. We also used 2, 3, 5-triphenyltetrazolium chloride (TTC) staining to assess the cerebral infarct volume. Then, we employed bioinformatics analysis and luciferase reporter assay to identify and validate the target molecule of miR-130a, PTEN. ResultsOur findings indicated that miR-130a expression was lower in PC12 cells after OGDR (oxygen-glucose deprivation/reperfusion) and in rats after MCAO (middle cerebral artery occlusion). Moreover, ectopic-expression of miR-130a can significantly improve cell survival rate and reduce cell apoptosis and ROS production in PC12 cells after OGDR. In addition, re-expression of miR-130a yielded an obvious reduction in MCAO-induced infarct volume and neurological deficits in rats. Bioinformatics analysis revealed that PTEN was a miR-130a target and could overturn the effect of miR-130a on cerebral ischemia, both in vivo and in vitro. Therefore, we set out to further investigate the PTEN-affected PI3K/AKT pathway and found that upregulation of miR-130a activated the PI3K/AKT pathway. ConclusionsOur data demonstrated that miR-130a prevented cerebral I/R damage by mediating the PTEN/PI3K/AKT axis. These preliminarily findings furthered our understanding of this mechanism and identified new potential therapeutic targets for ischemic stroke.

Highlights

Stroke has been, and continues to be, an important cause of death and disability in many industrialized nations
The results revealed that oxygen-glucose deprivation/Reperfusion (OGDR) treatment significantly inhibited the survival of PC12 cells when compared with the normoxic control, while overexpression of miR-130a resulted in an obvious increase in PC12 cell survival when compared with control mimic group (Fig. 1E)
The results of cell apoptosis analysis indicated that the apoptosis rate of PC12 cells increased in the OGDR group, while there was a decrease in the rate of apoptosis of PC12 cells in the miR-130a mimic group (Fig. 1F)

Summary

Introduction

Continues to be, an important cause of death and disability in many industrialized nations. Cerebral ischemic injury is a complicated pathological and physiological process, involving energy failure, acidosis, calcium overload, excitatory amino acid toxicity, mitochondrial damage, oxidative stress, and the inflammatory response. These mechanisms of damage result in cell necrosis or apoptosis [3,4,5]. Results: Our findings indicated that miR-130a expression was lower in PC12 cells after OGDR (oxygen-glucose deprivation/reperfusion) and in rats after MCAO (middle cerebral artery occlusion). Conclusions: Our data demonstrated that miR-130a prevented cerebral I/R damage by mediating the PTEN/ PI3K/AKT axis These preliminarily findings furthered our understanding of this mechanism and identified new potential therapeutic targets for ischemic stroke

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Conclusion