nr2a反义寡核苷酸对脑缺血-再灌注大鼠海马nr2a及其mrna表达的影响

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1、NR2A反义寡核苷酸对脑缺血/再灌注大鼠海马NR2A及其mRNA表达的影响：张旭，刘志安，滕大才，高伟，徐铁军【摘要】目的研究NMDA受体亚单位2A(NR2A)反义寡核苷酸对短暂性脑缺血/再灌注大鼠海马CA1区NR2A及其mRNA表达的影响，探讨其在脑缺血/再灌注诱导的大鼠海马神经元损伤中的可能作用机制。方法健康雄性SD大鼠随机分为正常对照组、假手术对照组和缺血/再灌注组。经生理盐水、错义寡核苷酸和反义寡核苷酸预处理后，以四血管阻断法建立短暂性前脑缺血(15min)再灌注(24h、48h和72h)动物模型。在确定

2、的时间点进行灌注固定、取材、石蜡包埋和组织切片(片厚8μm)，然后行原位杂交和免疫组织化学染色。结果短暂性脑缺血/再灌注后，大鼠海马CA1区NR2AmRNA的表达显著增加，与对照组相比差异有统计学意义(P<0.05);NR2A反义寡核苷酸能显著地抑制这种表达的增加(P<0.05)。短暂性脑缺血/再灌注后，大鼠海马CA1区NR2A的蛋白表达显著降低，与对照组相比差异有统计学意义(P<0.05);经NR2A反义寡核苷酸预处理后，能够进一步增强NR2A蛋白表达的降低，与对照组相比差异有统计学意义(P&

3、lt;0.05)。结论短暂性脑缺血/再灌注后，在大鼠海马CA1区存在转录增强而翻译抑制现象。NR2A反义寡核苷酸能特异性地抑制NR2AmRNA表达的增加，同时能够增强NR2A蛋白表达的降低。NR2A反义寡核苷酸的这种作用很可能与缺血后的翻译抑制以及海马CA1区选择性易损伤相关联。【关键词】脑缺血;NR2A反义寡核苷酸;NR2A;NR2AmRNA;翻译抑制　　Abstract:ObjectiveToinvestigatetheeffectofNMDAreceptorsubunit2A(NR2A)antisenseo

4、ligodeoxynucleotidesontheexpressionofNR2AanditsmRNAinrathippocampalCA1regionfolloia/reperfusion,andtofurtherprobeintothepossiblemechanismofneuronalcellinjuryinducedbycerebralischemiaandreperfusioninrathippocampus.MethodsHealthymaleSDratsizedintonormalcontrolg

5、roup,shamoperationcontrolgroupandischemia/reperfusiongroup.Folloentissenseoligonucleotideandantisenseoligonucleotide,thefour-vesselocclusionmethodployedtoestablishtransientforebrainischemia(15min)andreperfusion(24h,48hand72h)animalmodels.Indeterminingtime,the

6、ratsplepreparation,paraffin-embeddingandtissuesections(8μmofslicethickness).Theslicesmunohistochemicalstaining.ResultsFolloia/reperfusion,theexpressionofNR2AmRNAsignificantlyincreasedinCA1subfieldofrathippocampus(P<0.05)andtheNR2Aantisenseoligonucleotidesc

7、ouldsignificantlyinhibittheincreaseofthisexpression(P<0.05).Subsequenttotransientcerebralischemia/reperfusion,theexpressionofNR2AproteinsignificantlydecreasedinCA1subfieldofrathippocampus(P<0.05)andpretreatmentia/reperfusion,thereenonoftranscriptionenha

8、ncementandtranslationinhibitioninCA1regionofrathippocampus.NR2AantisenseoligonucleotidescouldspecificallyinhibittheincreaseinNR2AmRNAexpression,andcouldsimultaneouslystrengthenthereductio