Silencing LncRNA NEAT1 Reduces Neuronal Damage Through MiR-193a-3p To Regulate MMP14 In Cerebral Ischemia-reperfusion Injury

Background:Ischemic stroke is one of the main causes of severe disability and death in the world.The brain injury caused by ischemic stroke is mainly caused by a series of neuropathological diseases,including hypoxia,oxidative stress,edema formation,inflammatory reaction,etc.Several clinical practices revealed that some patients emerge with severe brain dysfunction rather than functional recovery when the blood reperfusion after cerebral ischemia.This clinical phenomenon is called cerebral ischemia-reperfusion injury(CIRI),which seriously affects the prognosis of patients and brings great challenges to the treatment of stroke.Therefore,the identification of cellular and molecular processes related to the pathogenesis and progression of CIRI has become a long-term orientation.Long non-coding RNAs(lncRNAs)are a class of non protein coding RNAs containing more than 200 nucleotides,which are involved in a series of biological process regulations,such as chromatin modification,transcriptional regulation,intranuclear transport,etc.Lnc RNA NEAT1 interacts with intracellular regulatory factors and plays an important role in the pathological process of cardiovascular and cerebrovascular diseases.Objective:To explore the mechanism of lncRNA NEAT1/mir-193a-3p /MMP14 axis in cerebral ischemia-reperfusion injury,we establish the oxygen and glucose deprivation/reoxygenation and middle cerebral artery occlusion/reperfusion models.Methods:1.The primary cortical neurons were treated with oxygen-glucose deprivation/reoxygenation(OGD/R)to construct the OGD/R model in vitro,and the rats in MCAO/R(middle cerebellar artery occlusion/reoxygenation)group and sham operation group were surgically constructed.The expression of lncRNA NEAT1 in cerebral ischemia-reperfusion injury model in vivo and in vitro were observed.Subsequently,transfection of lncRNA NEAT1 interfered with lentivirus into primary neuronal cells,and the changes in apoptosis and cytotoxicity of primary neuronal cells treated with OGD/R were observed.2.The competitive endogenous RNA(ce RNA)mi R-193a-3p and target gene MMP14 were analyzed and predicted by the bioinformatics website Starbase and micro RNA.org.Lentivirus si RNA NEAT1,plasmid MMP14,si RNA MMP14,and mir-193a-3p inhibitor were injected into the primary neuron cell model,respectively.The changes in apoptosis and cytotoxicity of primary neuronal cells treated with OGD /R were observed.3.Lv-NEAT1-sh RNA lentivirus was injected into the lateral ventricle of rats.The MCAO/R model of rats was constructed after 7 days.The neurological function,cerebral infarction volume,and neuronal apoptosis of rats were evaluated by Bederson score,TTC staining,HE staining,and TUNEL staining after MCAO/R.Results:1.It was found that the expression of lncRNA NEAT1 was significantly upregulated in cortical brain tissue of rats and primary neuronal cells induced by OGD/R;Interfering with the expression of lncRNA NEAT1 can significantly reduce the apoptosis of primary neuronal cells,the release of LDH and the expression of apoptosis-related proteins in primary neuronal cells treated by OGD/R.2.The protective effect of ischemic injury in neuronal cells by interfering si RNA NEAT1 could be significantly inhibited by si RNA NEAT1 and mir-193a-3p inhibitor co-transfection;The protective effect of ischemic injury in neuronal cells by interfering si RNA NEAT1 was not changed after interfering both si RNA NEAT1 and si RNA MMP14;Overexpression of MMP14 not only increased neuronal apoptosis induced by OGD/R treatment but also inhibited the protective effect of si RNA NEAT1.3.Compared with the NC group,the volume of cerebral infarction and neurological deficit score in the MCAO/R group were increased significantly,and the cortical neurons nuclear were deep staining,pyknosis of cortical neurons and vacuoles increased;The apoptosis rate of cortical neurons increased significantly.In the LVNEAT1-sh RNA group,the infarct volume was significantly reduced and the neurological deficit score was significantly improved;The hyperchromatism,nuclear pyknosis,and vacuole formation of cortical neurons were significantly reduced,and the apoptosis rate of cortical neurons was also significantly reduced.Conclusions:The expression of mi R-193a-3p was significantly down-regulated,while NEAT1 and MMP14 were significantly up-regulated in the primary neuronal OGD/R and MCAO/R model,suggesting that lncRNA NEAT1,mi R-193a-3p,and MMP14 play an important role in cerebral ischemia-reperfusion injury.The results of the mechanism study show that lncRNA NEAT1 regulates the expression of MMP14 by adsorbing mir-193a-3p,and promotes neuronal injury induced by cerebral ischemia-reperfusion injury.Our study reveals the lncRNA NEAT1/mi R-193a-3p/MMP14 functional axis in CIRI,which provides a new potential therapeutic target for CIRI.