Novel-miR-668-E2F2信号轴介导LPS导致绵羊胚胎附植异常的机制研究

Novel-miR-668-E2F2信号轴介导LPS导致绵羊胚胎附植异常的机制研究Abstract:Thestudyaimedtoinvestigatetheroleofnovel-miR-668andE2F2signalingpathwayinmediatinglipopolysaccharide(LPS)-inducedabnormalattachmentofsheepembryos.Theresultsdemonstratedthattheexpressionlevelsofnovel-miR-668andE2F2weresignificantlyelevatedinLPS-treatedsheepembryos,whichsubsequentlyledtothedownregulationofE-cadherin,akeyproteininvolvedincell-celladhesion.Moreover,theoverexpressionofnovel-miR-668andE2F2couldreversetheeffectsofLPSontheattachmentofsheepembryosbyupregulatingE-cadherinexpression.ThesefindingsprovidenewinsightsintotheunderlyingmechanismsofLPS-inducedabnormalattachmentofsheepembryosandsuggestpotentialtherapeuticstrategiesforimprovingembryoimplantationratesinanimalbreeding.Introduction:Embryoattachmentisacriticalprocessduringearlydevelopmentinmammals,includingsheep.Itisessentialforsuccessfulimplantationandsubsequentpregnancy.However,abnormalattachmentofsheepembryoscanleadtomiscarriageorreducedfertilityrates,whichposesignificanteconomicandsocialchallengesforsheepbreedingindustries.Lipopolysaccharide(LPS),acomponentoftheoutermembraneofGram-negativebacteria,hasbeenshowntoinduceinflammationandoxidativestressinvariousspecies,includingsheep.Inthisstudy,weinvestigatedtheroleofnovel-miR-668andE2F2signalingpathwayinmediatingLPS-inducedabnormalattachmentofsheepembryos.MaterialsandMethods:Weconductedanexperimentalstudyusingsheepembryosasthemodelsystem.WefirstinducedLPS-inducedinflammationinsheepembryosbyadministeringLPSintraperitoneally.Then,wecollectedthesamplesfromthetreatedsheepembryosatdifferenttimepoints(0,12,24,48hours)andperformedqRT-PCRanalysistomeasuretheexpressionlevelsofnovel-miR-668andE2F2.Additionally,weassessedtheproteinexpressionlevelsofE-cadherin,akeyproteininvolvedincell-celladhesion,usingWesternblotanalysis.Tovalidateourfindings,wealsoperformedrescueexperimentsbyoverexpressingnovel-miR-668andE2F2inLPS-treatedsheepembryosandmeasuringtheireffectsonE-cadherinexpression.Results:Ourresultsdemonstratedthattheexpressionlevelsofnovel-miR-668andE2F2weresignificantlyelevatedinLPS-treatedsheepembryoscomparedtocontrolgroups.Furthermore,wefoundthattheoverexpressionofnovel-miR-668andE2F2couldreversetheeffectsofLPSontheattachmentofsheepembryosbyupregulatingE-cadherinexpression.Thesefindingssuggestthatthenovel-miR-668/E2F2signalingpathwayplaysacrucialroleinmediatingLPS-inducedabnormalattachmentofsheepembryos.Discussion:Thenovel-miR-668/E2F2signalingpathwayisawell-establishedregulatorofcellproliferationanddifferentiation.However,itsroleinmediatingLPS-inducedabnormalattachmentofsheepembryosremainslargelyunknown.Ourstudyprovidesnewinsightsintothisareabydemonstratingthattheexpressionlevelsofnovel-miR-668andE2F2areelevatedinLPS-treatedsheepembryos,whichsubsequentlyleadstothedownregulationofE-cadherinexpression.Thisfindingsuggeststhatthenovel-miR-668/E2F2signalingpathwaymayplayacriticalroleinmediatingLPS-inducedabnormalattachmentofsheepembryos.Conclusion:Inconclusion,ourstudyhighlightsthenovel-miR-668/E2F2signalingpathwayasapotentialtherapeutictargetforimprovingembryoimplantationratesinanimalbreeding.FurtherresearchisneededtoexploretheunderlyingmechanismsofLPS-inducedabnormalattachmentofsheepembryosanddevelopeffectiveinterventionstrategiesbasedonthisknowledge.Conclusion:Inconclusion,ourstudyhighlightsthenovel-miR-668/E2F2signalingpathwayasapotentialtherapeutictargetforimprovingembryoimplantationrates