mirnalet-7c-5p对缺血性脑损伤的保护作用与其机制分析

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1、中文摘要miRNAlet-7c-5p对缺血性脑损伤的保护作用及其机制研究caspase3种子序列突变的突变型HEK293细胞相比，let-7c-5p过表达可以显著降低共转染野生型caspase3HEK293细胞的荧光强度。同时，过表达let-7c-5p可以显著抑制由LPS诱导的总caspase3和剪切的caspase3的表达。结论:过表达let-7c-5p对缺血再灌注损伤具有保护作用，直接靶向caspase3从而抑制小胶质细胞的激活可能是这种保护作用的机制之一。关键词：miRNA；let-7c-5p；脑缺

2、血；小胶质细胞作者：倪敬书指导教师：镇学初IImiRNAlet-7c-5p对缺血性脑损伤的保护作用及其机制研究英文摘要MicroRNAlet-7c-5pprotectsagainstcerebralischemiainjuryviamechanismsinvolvingtheinhibitionofmicrogliaactivationAbstractObjective:WeobservedthatLet-7c-5p,ahighlyconservedmiRNA,wasdownregulatedinthep

3、lasmaofpatientswithischemicstroke.Here,weinvestigatedtheeffectsoflet-7c-5ponischemicbraindamageandpotentialmechanisms.Methods:Mousecerebralischemiawasinducedbythemiddlecerebralarteryocclusion(MCAO).PrimarymicrogliaandBV2microglialcellswereactivatedinvitro

4、byLPS,oxygen-glucosedeprivation(OGD)orOGD-conditionedneuronalbuffer.Acutecerebralinfarctionwasdeterminedby2,3,5-triphenyltertrazoliumchloridestaining.Immunostainingwasperformedtocharacterizetheactivationofmicroglia.Theexpressionofinflammatorymediatorsinth

5、eischemicbrainandactivatedmicrogliawasanalyzedwithqPCRandwesternblotting.Luciferasereporterassaywasperformedtoidentifythesiteinthe3’un-translatedregionofthetargetmRNAthatwastargetedbylet-7c-5p.Results:let-7c-5plevelsweresignificantlydecreasedintheplasmaof

6、patientswithischemicstroke,aswellasinmouseplasma,ipsilateralcortexandstriatumat6and24hoursfollowingMCAO.Overexpressionoflet-7c-5pbyICVinjectionofmiRNAmimicsdecreasedinfarctionandmicrogliaactivation,asevidencedbythefactthatlet-7c-5poverexpressionattenuated

7、expressionofpro-inflammatorymediatorsandramifiedIba1immunoactivityintheischemicbrain.Invitrostudyfurtherrevealedthatlet-7c-5pwasreducedintheactivatedmicrogliaandlet-7c-5poverexpressionremarkablyinhibitedactivation-inducedexpressionofseveralpro-inflammator

8、ymediatorsinmicroglia.Luciferasereporterassaysuggestthatlet-7c-5pdirectlytargetthe3’-untranslatedregionofcaspase3mRNAtoreducethemRNAlevelsofcaspase3inmicroglia.Conclusions:Let-7c-5poverexpressionprotectsagainstische