Effects and mechanisms of LINC00657 on oxidative glucose deprivation-induced injury in mouse hippocampal neurons
Objective To investigate the effects and mechanisms of LINC00657 on oxidative glu-cose deprivation(OGD)-induced injury in mouse hippocampal neurons.Methods Mouse hippocam-pal neuron cell line HT22 was given OGD treatment to establish an injury model,with normally cul-tured HT22 cells as controls.The si-NC,si-LINC00657,microRNA(miR)-NC,and miR-224-3p mimics were transfected into HT22 cells,followed by OGD treatment.Co-transfection of si-LINC00657 and anti-miR-NC,or co-transfection of si-LINC00657 and anti-miR-224-3p,was performed in HT22 cells before OGD treatment.Real-time quantitative reverse transcription-polymerase chain reaction(qRT-PCR)was used to detect the relative expression levels of LINC00657 and miR-224-3p.CCK-8 assay and flow cytometry were used to detect cell viability and apoptosis rate,respectively.Kits were used to detect the activities of lactate dehydrogenase(LDH),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),and the level of malondialdehyde(MDA).Dual-luciferase reporter gene assay was used to detect the effect of miR-224-3p overexpression on the luciferase activity of wild-type LINC00657 vector(WT-LINC00657)and mutant LINC00657 vector(MUT-LINC00657).Results Compared with controls,the expression of LINC00657 was upregulated and the expression of miR-224-3p was downregulated in OGD-induced HT22 cells(P＜0.05).Compared with trans-fection of si-NC or miR-NC,transfection of si-LINC00657 or miR-224-3p mimics resulted in in-creased cell viability,SOD activity,and GSH-Px activity,as well as decreased apoptosis rate,LDH activity,and MDA level(P＜0.05).Overexpression of miR-224-3p reduced the luciferase activity of WT-LINC00657(P＜0.05).Compared with cells co-transfected with si-LINC00657 and anti-miR-NC,cells co-transfected with si-LINC00657 and anti-miR-224-3p showed decreased cell viabili-ty,increased apoptosis rate,increased LDH activity and MDA level,and decreased SOD and GSH-Px activities(P＜0.05).Conclusion Interference with LINC00657 can promote cell proliferation,inhibit apoptosis and oxidative stress response by upregulating miR-224-3p,thereby alleviating OGD-induced injury in mouse hippocampal neurons.
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