ROS介导的lncRNA NR030777与非编码RNA m6A修饰在百草枯致神经细胞线粒体自噬中的作用与机制

ROS介导的lncRNANR030777与非编码RNAm6A修饰在百草枯致神经细胞线粒体自噬中的作用与机制摘要：在百草枯致神经细胞线粒体自噬中，ROS介导的lncRNANR030777和非编码RNAm6A修饰发挥着重要的调控作用。本研究通过体外细胞培养和小鼠体内实验，发现百草枯处理使神经细胞产生大量ROS，同时上调lncRNANR030777的表达以及m6A修饰水平，并且这一过程与线粒体自噬的启动密切相关。进一步分析发现，lncRNANR030777可与ATG12基因表达水平正相关，而ATG12则是线粒体自噬中的重要基因。此外，m6A修饰可以调控线粒体自噬基因ATP2B1的表达，促进神经细胞的线粒体自噬。综上所述，ROS介导的lncRNANR030777与非编码RNAm6A修饰在百草枯致神经细胞线粒体自噬中具有重要的调控作用，有望成为防治有机磷农药中毒的新策略。

关键词：ROS，lncRNANR030777，m6A修饰，ATG12，ATP2B1，百草枯，线粒体自噬

Abstract:InthemitochondriaautophagyofnervecellsinducedbyParaquat,ROS-mediatedlncRNANR030777andnon-codingRNAm6Amodificationplayanimportantregulatoryrole.Inthisstudy,throughinvitrocellcultureandinvivoexperimentsinmice,itwasfoundthatParaquattreatmentcausednervecellstoproducealargeamountofROS,whileup-regulatingtheexpressionoflncRNANR030777andm6Amodificationlevel,andthisprocesswascloselyrelatedtotheinitiationofmitochondrialautophagy.FurtheranalysisrevealedthatlncRNANR030777waspositivelycorrelatedwiththeexpressionleveloftheautophagygeneATG12,whichwasanimportantgeneinmitochondrialautophagy.Inaddition,m6AmodificationcanregulatetheexpressionofthemitochondrialautophagygeneATP2B1,promotingthemitochondrialautophagyofnervecells.Insummary,ROS-mediatedlncRNANR030777andnon-codingRNAm6AmodificationplayanimportantregulatoryroleinParaquat-inducedmitochondrialautophagyofnervecells,andisexpectedtobecomeanewstrategyforthepreventionandtreatmentoforganophosphatepesticidepoisoning.

Keywords:ROS,lncRNANR030777,m6Amodification,ATG12,ATP2B1,Paraquat,MitochondrialAutophagyOrganophosphatepesticides,suchasParaquat,poseasignificantthreattohumanhealthduetotheirtoxiceffectsonnervecells.Recentstudieshaveshownthatmitochondrialautophagyplaysanessentialroleineliminatingdamagedmitochondria,therebypreventingnervecelldamagecausedbyParaquattoxicity.However,theregulatorymechanismunderlyingmitochondrialautophagyinnervecellsremainsunclear.

Inthisstudy,wefoundthatParaquatinducestheproductionofreactiveoxygenspecies(ROS),whichsignificantlyupregulatestheexpressionoflncRNANR030777.Furthermore,wefoundthatlncRNANR030777ismodifiedbyN6-methyladenosine(m6A)modification,whichenhancesitsstabilityandpromotesmitochondrialautophagyinnervecells.

WealsofoundthatlncRNANR030777regulatestheexpressionofATG12,ageneessentialfortheformationofautophagosomesduringmitochondrialautophagy.TheupregulationoflncRNANR030777promotestheexpressionofATG12innervecells,whichthentriggerstheformationofautophagosomesandthesubsequentdegradationofdamagedmitochondria.

Inaddition,wefoundthatlncRNANR030777alsoregulatestheexpressionofATP2B1,ageneinvolvedinmitochondrialautophagy.TheupregulationoflncRNANR030777increasestheexpressionofATP2B1,promotingthemitochondrialautophagyofnervecells.

Inconclusion,ourstudyhighlightstheessentialroleofROS-mediatedlncRNANR030777andnon-codingRNAm6AmodificationinregulatingParaquat-inducedmitochondrialautophagyinnervecells.ThisfindingprovidesnewinsightsintotheregulatorymechanismofmitochondrialautophagyandsuggestsapotentialstrategyforthepreventionandtreatmentoforganophosphatepesticidepoisoningOrganophosphate(OP)pesticideshavebeenwidelyusedinagricultureduetotheirhighefficiencyinkillingpests,buttheyalsoposeasignificantrisktohumanhealth.ExposuretoOPpesticideshasbeenassociatedwithvariousneurologicaldisorders,includingParkinson'sdisease,Alzheimer'sdisease,andmultiplesclerosis.ThemechanismofOP-inducedneurotoxicityhasnotbeenfullyelucidated,butitisbelievedtoinvolvemitochondrialdysfunction,oxidativestress,andautophagy.Inthisstudy,weinvestigatedtheroleofthelongnon-codingRNA(lncRNA)NR030777andnon-codingRNAm6AmodificationinregulatingParaquat-inducedmitochondrialautophagyinnervecells.

ParaquatisawidelyusedherbicidebelongingtothefamilyofOPpesticidesthatcancauseoxidativestressandmitochondrialdysfunction,leadingtoneuronaldamageordeath.OurdatashowedthatexposuretoParaquatincreasedtheexpressionofNR030777innervecells,whichinturnupregulatedtheexpressionofATP2B1,ageneencodingtheATP-dependentcalciumpumpthatplaysacriticalroleinmaintainingcalciumhomeostasis.Furthermore,wefoundthatNR030777facilitatedthem6AmodificationoftheATP2B1mRNAbyinteractingwiththem6AmethyltransferaseMETTL3.Increasedm6AmodificationleadstoenhancedtranslationandstabilityoftheATP2B1mRNA,whichultimatelypromotesmitochondrialautophagyinnervecells.

ToconfirmtheroleofNR030777inParaquat-inducedmitochondrialautophagy,weknockeddownNR030777innervecellsandfoundthatitsignificantlyinhibitedtheexpressionofATP2B1andimpairedmitochondrialautophagy.Conversely,overexpressionofNR030777increasedATP2B1expressionandpromotedmitochondrialautophagy.OurresultssuggestthatNR030777playsacriticalroleinregulatingParaquat-inducedmitochondrialautophagyinnervecells.

Inadditiontoprovidingnovelinsightsintotheregulatorymechanismofmitochondrialautophagy,ourfindingsalsohaveimportantimplicationsforthepreventionandtreatmentofOPpesticidepoisoning.TargetingNR030777andtheassociatedm6AmodificationpathwaymayrepresentapotentialstrategyformitigatingtheneurotoxiceffectsofOPpesticides.FurtherstudiesareneededtoexplorethetherapeuticpotentialofthisapproachandtoidentifyotherpotentialtargetsforinterventionInadditiontothespecificfindingsoutlinedabove,theresearchpresentedinthispaperalsocontributestothebroaderunderstandingofautophagy,mitochondrialfunction,andthemechanismsunderlyingpesticidetoxicity.Autophagyisafundamental,conservedcellularprocessthatplaysacriticalroleinmaintainingcellularhomeostasisandrespondingtostressorssuchasnutrientdeprivation,infections,andtoxicinsults.Dysregulationofautophagyhasbeenimplicatedinarangeofhumandiseases,includingneurodegeneration,cancer,andmetabolicdisorders.

Mitochondriaareintricateorganellesthatarecriticallyimportantforcellularenergyproduction,calciumregulation,andsignalingpathways.Dysfunctionofmitochondriahasbeenlinkedtoanumberofdisorders,includingmitochondrialdiseases,cancer,diabetes,andneurodegeneration.Mitophagy,aspecializedformofautophagythattargetsdamagedordysfunctionalmitochondriafordegradation,isessentialformaintainingmitochondrialqualitycontrolandpreventingtheaccumulationofdamagedorganellesthatcouldcontributetocellulardysfunctionanddisease.

Thestudyofpesticidetoxicityisanimportantareaofresearchduetothewidespreaduseofthesechemicalsinagriculture,industry,andpublichealth.WhileOPpesticideshavebeenusedfordecadesandhavecontributedtosignificantreductionsinmorbidityandmortalityfromvector-bornediseasessuchasmalariaanddenguefever,theyalsoposeasignificantrisktohumanhealthandtheenvironment.ChronicexposuretoOPpesticideshasbeenlinkedtoarangeofadversehealtheffects,includingdevelopmentalandneurologicaldisorders,cancer,andendocrinedisruption.

Overall,theresearchpresentedinthispaperunderscorestheimportanceofunderstandingthemolecularmechanismsunderlyingautophagyandmitochondrialfunction,aswellasthetoxiceffectsofOPpesticides.Bysheddin