MicroRNA-31激活ROCK1介導(dǎo)的PI3K-AKT信號(hào)通路改善LPS誘導(dǎo)的內(nèi)皮細(xì)胞炎癥和通透性研究

MicroRNA-31激活ROCK1介導(dǎo)的PI3K-AKT信號(hào)通路改善LPS誘導(dǎo)的內(nèi)皮細(xì)胞炎癥和通透性研究摘要

背景：內(nèi)皮細(xì)胞炎癥和通透性是炎癥反應(yīng)和疾病發(fā)展的關(guān)鍵環(huán)節(jié)。微小RNA（miRNA）作為一個(gè)小分子RNA參與細(xì)胞信號(hào)傳遞、基因調(diào)控和蛋白翻譯等過(guò)程，在多種疾病的發(fā)生和進(jìn)展中發(fā)揮著重要作用。ROCK1是一個(gè)關(guān)鍵的細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)因子，與PI3K/AKT信號(hào)通路緊密相關(guān)，參與細(xì)胞的運(yùn)動(dòng)、增殖和凋亡等生物學(xué)過(guò)程。

方法：本研究選取LPS刺激的人體內(nèi)皮細(xì)胞為研究對(duì)象，應(yīng)用定量PCR、Westernblot和免疫熒光等方法檢測(cè)細(xì)胞中miRNA-31、ROCK1、PI3K/AKT等信號(hào)通路相關(guān)因子的表達(dá)情況，并觀察在miRNA-31激活ROCK1介導(dǎo)下對(duì)內(nèi)皮細(xì)胞炎癥和旁路引起的通透性的影響。

結(jié)果：實(shí)驗(yàn)結(jié)果顯示，LPS刺激后，內(nèi)皮細(xì)胞中miRNA-31的表達(dá)下調(diào)，ROCK1和PI3K/AKT的表達(dá)上調(diào)；miRNA-31激活ROCK1可促進(jìn)PI3K/AKT信號(hào)通路的激活，減輕內(nèi)皮細(xì)胞的炎癥反應(yīng)和通透性的損傷；而抑制ROCK1則弱化內(nèi)皮細(xì)胞對(duì)炎癥和通透性的應(yīng)答能力。

結(jié)論：miRNA-31通過(guò)激活ROCK1介導(dǎo)的PI3K/AKT信號(hào)通路，改善LPS誘導(dǎo)的內(nèi)皮細(xì)胞炎癥和通透性。本研究結(jié)果提示miRNA-31-ROCK1-PI3K/AKT信號(hào)通路可能成為內(nèi)皮細(xì)胞保護(hù)和治療炎癥相關(guān)疾病的新靶點(diǎn)。

關(guān)鍵詞：內(nèi)皮細(xì)胞炎癥；內(nèi)皮細(xì)胞通透性；微小RNA-31；ROCK1；PI3K/AKT信號(hào)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