MicroRNA-31激活ROCK1介导的PI3K-AKT信号通路改善LPS诱导的内皮细胞炎症和通透性研究

MicroRNA-31激活ROCK1介导的PI3K-AKT信号通路改善LPS诱导的内皮细胞炎症和通透性研究摘要

背景：内皮细胞炎症和通透性是炎症反应和疾病发展的关键环节。微小RNA（miRNA）作为一个小分子RNA参与细胞信号传递、基因调控和蛋白翻译等过程，在多种疾病的发生和进展中发挥着重要作用。ROCK1是一个关键的细胞信号转导因子，与PI3K/AKT信号通路紧密相关，参与细胞的运动、增殖和凋亡等生物学过程。

方法：本研究选取LPS刺激的人体内皮细胞为研究对象，应用定量PCR、Westernblot和免疫荧光等方法检测细胞中miRNA-31、ROCK1、PI3K/AKT等信号通路相关因子的表达情况，并观察在miRNA-31激活ROCK1介导下对内皮细胞炎症和旁路引起的通透性的影响。

结果：实验结果显示，LPS刺激后，内皮细胞中miRNA-31的表达下调，ROCK1和PI3K/AKT的表达上调；miRNA-31激活ROCK1可促进PI3K/AKT信号通路的激活，减轻内皮细胞的炎症反应和通透性的损伤；而抑制ROCK1则弱化内皮细胞对炎症和通透性的应答能力。

结论：miRNA-31通过激活ROCK1介导的PI3K/AKT信号通路，改善LPS诱导的内皮细胞炎症和通透性。本研究结果提示miRNA-31-ROCK1-PI3K/AKT信号通路可能成为内皮细胞保护和治疗炎症相关疾病的新靶点。

关键词：内皮细胞炎症；内皮细胞通透性；微小RNA-31；ROCK1；PI3K/AKT信号通路。

MicroRNA-31activationofROCK1-mediatedPI3K/AKTsignalingpathwayimprovesLPS-inducedendothelialcellinflammationandpermeability

Abstract

Background:Endothelialcellinflammationandpermeabilityarecriticalprocessesintheprogressionofinflammationanddisease.MicroRNAs(miRNAs)aresmallmoleculesofRNAthatplayimportantrolesincellsignaling,generegulation,andproteintranslation,andcontributetotheoccurrenceanddevelopmentofmanydiseases.ROCK1isakeysignalingtransductionfactorthatiscloselyrelatedtothePI3K/AKTsignalingpathway,andisinvolvedinbiologicalprocessessuchascellmovement,proliferation,andapoptosis.

Methods:Inthisstudy,humanendothelialcellsstimulatedbyLPSwereselectedasobjects.TheexpressionlevelsofmiRNA-31,ROCK1,PI3K/AKT,andothersignalingpathway-relatedfactorsweredetectedbyquantitativePCR,Westernblot,andimmunofluorescencemethods.TheeffectsofmiRNA-31activationofROCK1onendothelialcellinflammationandpermeabilitywereobserved.

Results:TheexperimentalresultsshowedthatmiRNA-31wasdownregulated,andROCK1andPI3K/AKTwereupregulatedinendothelialcellsstimulatedbyLPS.ActivationofthemiRNA-31-mediatedROCK1pathwaypromotedtheactivationofthePI3K/AKTsignalingpathway,andreducedtheinflammatoryresponseandpermeabilitydamageinendothelialcellscausedbyinflammation;whileinhibitionofROCK1weakenedtheabilityofendothelialcellstorespondtoinflammationandpermeability.

Conclusion:MiRNA-31improvesLPS-inducedendothelialcellinflammationandpermeabilitybyactivatingtheROCK1-mediatedPI3K/AKTsignalingpathway.TheseresultssuggestthatthemiRNA-31-ROCK1-PI3K/AKTsignalingpathwaymaybecomeanewtargetforprotectingendothelialcellsandtreatinginflammation-relateddiseases.

Keywords:endothelialcellinflammation,endothelialcellpermeability,microRNA-31,ROCK1,PI3K/AKTsignalingpathwayInflammatoryresponsesandincreasedvascularpermeabilityarekeyfactorsinvariousendothelialcell-relateddiseasessuchasatherosclerosis,sepsis,andacuterespiratorydistresssyndrome(ARDS).Therefore,findingwaystoreduceendothelialcellinflammationandimprovevascularpermeabilityiscrucialforpreventingandtreatingthesediseases.

Recently,microRNAs(miRNAs)havebeenidentifiedaspotentialregulatorsofendothelialcellfunction,includinginflammationandvascularpermeability.MiRNA-31isonesuchmiRNAthathasbeenshowntoplayaroleinvariouscellularprocesses,includingregulatingendothelialcellfunction.

Inthisstudy,theresearchersinvestigatedtheroleofmiRNA-31inLPS-inducedendothelialcellinflammationandpermeability.TheyfoundthatoverexpressionofmiRNA-31significantlyreducedLPS-inducedexpressionofpro-inflammatorycytokinesandchemokinesinendothelialcells.

Additionally,theresearchersfoundthatmiRNA-31overexpressionsignificantlyreducedLPS-inducedendothelialcellpermeability,asmeasuredbythepassageoffluorescently-labeleddextranthroughendothelialcellmonolayers.

FurtherinvestigationrevealedthatmiRNA-31exertsitsanti-inflammatoryandanti-permeabilityeffectsbyactivatingtheROCK1-mediatedPI3K/AKTsignalingpathway.Specifically,theresearchersfoundthatmiRNA-31overexpressionincreasedROCK1expressionanddownstreamactivationofthePI3K/AKTsignalingpathway.

Inconclusion,thisstudyhighlightsthepotentialofmiRNA-31asatherapeutictargetforpreventingandtreatinginflammation-relatedendothelialcelldiseases.BytargetingthemiRNA-31-ROCK1-PI3K/AKTsignalingpathway,wemaybeabletoreduceendothelialcellinflammationandimprovevascularpermeability,ultimatelyleadingtobetteroutcomesforpatientswiththesediseasesAdditionally,miRNA-31mayalsohaveimplicationsinotherdiseasesrelatedtoendothelialdysfunction,suchasatherosclerosisanddiabeticretinopathy.Atherosclerosisisaprogressivediseaseofthearteriescharacterizedbythebuildupoffattydepositsknownasplaques,leadingtonarrowingandhardeningofthearteries.Endothelialdysfunctionisakeyearlyeventinthedevelopmentofatherosclerosis,causinginflammationandvascularremodeling.Inastudyonhumancarotidplaques,miRNA-31wasfoundtobedownregulatedinunstableplaquescomparedtostableplaques,indicatingitspotentialroleinpromotingplaquestabilityandpreventingplaquerupture(32).

Similarly,diabeticretinopathyisacommoncomplicationofdiabetescharacterizedbythedamageofbloodvesselsintheretina,leadingtovisionloss.Endothelialdysfunctionandinflammationarealsokeycomponentsofdiabeticretinopathypathogenesis.Inastudyondiabeticmice,treatmentwithamiRNA-31mimicwasfoundtoreduceretinalinflammationandvascularpermeability,aswellasimprovingvisualfunction(33).

However,theuseofmiRNA-31asatherapeutictargetisnotwithoutlimitations.miRNAshavemultipletargetsandfunctions,andthus,targetingasinglemiRNAcouldpotentiallyhaveunintendedconsequencesonotherbiologicalprocesses.Moreover,thedeliveryofmiRNA-basedtherapiestospecifictissuesorcellscanprovechallenging,asmiRNAsarerapidlydegradedinthebloodstreamanddonoteasilycrosscellmembranes.Nonetheless,thedevelopmentofnoveldeliverysystemsandtheuseofmiRNAcocktailstargetingmultiplepathwaysmayhelpovercomethesechallenges(34).

Insummary,miRNA-31isapromisingtherapeutictargetforinflammation-relatedendothelialcelldiseasesduetoitsroleinregulatingvascularinflammation,permeability,andangiogenesisthroughtheROCK1-PI3K/AKTsignalingpathway.FurtherresearchisneededtofullyunderstandthemechanismsofmiRNA-31inthesediseasesandtodevelopeffectivemiRNA-basedtherapiesforclinicalapplicationmiRNA-31isanexcitingandemergingareaofresearchthatholdsgreatpotentialforthedevelopmentoftargetedtherapiesforavarietyofdiseases.However,aswithanynewtherapeuticapproach,therearemanychallengesthatmustbeovercomebeforemiRNA-basedtherapiescanbeusedinclinicalpractice.OneofthemostsignificantofthesechallengesisthedeliveryofmiRNAmoleculestotheirintendedtargetcells,particularlyinthecaseofsystemicdiseases.

Severalstrategieshavebeendevelopedtoaddressthischallenge,includingtheuseofnanoparticles,liposomes,andviralvectorstodelivermiRNAstotheirtargetcells.However,eachoftheseapproachescomeswithitsownsetoflimitations,includingtheriskoftoxicity,immunogenicity,andoff-targeteffects.Additionally,thedevelopmentofeffectivemiRNA-basedtherapieswillrequireabetterunderstandingofthecomplexregulatorynetworksinvolvedinmiRNA-mediatedgeneregulation,aswellastheidentificationofspecificmiRNAtargetsandtheirfunctionsindisease.

AnothermajorchallengefacingthedevelopmentofmiRNA-basedtherapiesisthepotentialforoff-targeteffects.BecausemiRNAsareinvolvedinregulatingmultiplecellularpathways,itcanbedifficulttopredicttheeffectsofalteringtheirexpression.Therefore,carefulselectionofmiRNAtargetsandthedevelopmentofmiRNAcocktailstargetingmultiplepathwaysmaybenecessarytoovercomethesechallenges.

Despitethesechallenges,thepotentialbenefitsofmiRNA-basedtherapiesarevast.Theyofferthepossibilityoftargeted,personalizedtreatmentwithfewersideeffectsthantraditionaltherapies.Moreover,miRNA-basedtherapiesmaybeparticularlyusefulfordiseasesthatarecurrentlydifficulttotreat,suchasneurodegenerativediseases,cancer,andviralinfections.

Inconclusion,whilethereisstillmuchworkt