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《nr2a反義寡核苷酸對(duì)腦缺血-再灌注大鼠海馬nr2a及其mrna表達(dá)的影響》由會(huì)員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在應(yīng)用文檔-天天文庫(kù)。
1、NR2A反義寡核苷酸對(duì)腦缺血/再灌注大鼠海馬NR2A及其mRNA表達(dá)的影響:張旭,劉志安,滕大才,高偉,徐鐵軍【摘要】目的研究NMDA受體亞單位2A(NR2A)反義寡核苷酸對(duì)短暫性腦缺血/再灌注大鼠海馬CA1區(qū)NR2A及其mRNA表達(dá)的影響,探討其在腦缺血/再灌注誘導(dǎo)的大鼠海馬神經(jīng)元損傷中的可能作用機(jī)制。方法健康雄性SD大鼠隨機(jī)分為正常對(duì)照組、假手術(shù)對(duì)照組和缺血/再灌注組。經(jīng)生理鹽水、錯(cuò)義寡核苷酸和反義寡核苷酸預(yù)處理后,以四血管阻斷法建立短暫性前腦缺血(15min)再灌注(24h、48h和72h)動(dòng)物模型。在確定
2、的時(shí)間點(diǎn)進(jìn)行灌注固定、取材、石蠟包埋和組織切片(片厚8μm),然后行原位雜交和免疫組織化學(xué)染色。結(jié)果短暫性腦缺血/再灌注后,大鼠海馬CA1區(qū)NR2AmRNA的表達(dá)顯著增加,與對(duì)照組相比差異有統(tǒng)計(jì)學(xué)意義(P<0.05);NR2A反義寡核苷酸能顯著地抑制這種表達(dá)的增加(P<0.05)。短暫性腦缺血/再灌注后,大鼠海馬CA1區(qū)NR2A的蛋白表達(dá)顯著降低,與對(duì)照組相比差異有統(tǒng)計(jì)學(xué)意義(P<0.05);經(jīng)NR2A反義寡核苷酸預(yù)處理后,能夠進(jìn)一步增強(qiáng)NR2A蛋白表達(dá)的降低,與對(duì)照組相比差異有統(tǒng)計(jì)學(xué)意義(P&
3、lt;0.05)。結(jié)論短暫性腦缺血/再灌注后,在大鼠海馬CA1區(qū)存在轉(zhuǎn)錄增強(qiáng)而翻譯抑制現(xiàn)象。NR2A反義寡核苷酸能特異性地抑制NR2AmRNA表達(dá)的增加,同時(shí)能夠增強(qiáng)NR2A蛋白表達(dá)的降低。NR2A反義寡核苷酸的這種作用很可能與缺血后的翻譯抑制以及海馬CA1區(qū)選擇性易損傷相關(guān)聯(lián)?!娟P(guān)鍵詞】腦缺血;NR2A反義寡核苷酸;NR2A;NR2AmRNA;翻譯抑制 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