黃芪甲苷對(duì)哮喘小鼠Th17-Treg細(xì)胞調(diào)控作用的研究黃芪甲苷對(duì)哮喘小鼠Th17/Treg細(xì)胞調(diào)控作用的研究

摘要：本研究旨在探討黃芪甲苷對(duì)哮喘小鼠Th17/Treg細(xì)胞的調(diào)控作用及其機(jī)制。采用小鼠哮喘模型，用流式細(xì)胞術(shù)、ELISA法和實(shí)時(shí)熒光定量PCR技術(shù)檢測(cè)小鼠肺組織細(xì)胞因子IL-17、IL-10的表達(dá)、Th17細(xì)胞(IL-17+)、Treg細(xì)胞(CD4+CD25+Foxp3+)比例及mRNA水平等指標(biāo)。結(jié)果表明：與模型組相比，黃芪甲苷可顯著降低小鼠肺組織IL-17水平，增加IL-10表達(dá)，下調(diào)Th17細(xì)胞比例和mRNA水平，同時(shí)提高Treg細(xì)胞數(shù)量及Foxp3mRNA水平(均P<0.05)。此外，黃芪甲苷可顯著降低小鼠肺組織MDA、ROS級(jí)別、增加SOD活力和GSH、T-AOC水平(均P<0.05)。上述結(jié)果表明，黃芪甲苷能夠通過(guò)免疫調(diào)節(jié)和抗氧化應(yīng)激作用，下調(diào)小鼠哮喘肺組織中Th17細(xì)胞比例和IL-17水平，提高Treg細(xì)胞比例、Foxp3mRNA水平和IL-10表達(dá)水平，從而發(fā)揮治療哮喘作用。

關(guān)鍵詞：黃芪甲苷，哮喘，Th17細(xì)胞，Treg細(xì)胞，IL-17，IL-10，免疫調(diào)節(jié)，抗氧化應(yīng)激

Abstract:ThisstudyaimstoinvestigatetheregulatoryeffectandmechanismofHuangqiJiaqinonTh17/Tregcellsinasthmaticmice.Themouseasthmamodelwasused,andflowcytometry,ELISA,andreal-timefluorescentquantitativePCRwereusedtodetecttheexpressionofIL-17,IL-10inlungtissue,theratioandmRNAlevelofTh17cells(IL-17+),Tregcells(CD4+CD25+Foxp3+),andotherindicators.Theresultsshowedthatcomparedwiththemodelgroup,HuangqiJiaqincouldsignificantlyreducethelevelofIL-17inlungtissue,increasetheexpressionofIL-10,down-regulatetheratioandmRNAlevelofTh17cells,andincreasethenumberofTregcellsandFoxp3mRNAlevel(allP<0.05).Inaddition,HuangqiJiaqincouldsignificantlyreducethelevelsofMDAandROSinlungtissue,increasetheactivityofSODandthelevelsofGSHandT-AOC(allP<0.05).TheaboveresultsindicatethatHuangqiJiaqincanregulateimmunityandrelieveoxidativestress,down-regulatetheratioandIL-17levelofTh17cellsinasthmaticmouselungtissue,increasetheratioofTregcells,Foxp3mRNAlevel,andIL-10expressionlevel,therebyexertingtherapeuticeffectsonasthma.

Keywords：HuangqiJiaqin,asthma,Th17cells,Tregcells,IL-17,IL-10,immuneregulation,anti-oxidativestressAsthmaisachronicinflammatorydiseaseoftheairways,characterizedbyvariableairflowobstructionandbronchialhyperresponsiveness.Itaffectsmillionsofpeopleworldwideandhasbecomeamajorpublichealthconcern.TraditionalChinesemedicinehasbeenwidelyusedasanalternativetherapyforasthma,andHuangqiJiaqinisaherbalformulathathasbeenreportedtoexhibittherapeuticeffectsonasthma.

Inthisstudy,weinvestigatedthepotentialmechanismofHuangqiJiaqininthetreatmentofasthmabyregulatingtheimmuneresponseandrelievingoxidativestress.OurresultsshowedthatHuangqiJiaqinsignificantlydecreasedtheratioandIL-17levelofTh17cellsinasthmaticmouselungtissuewhileincreasingtheratioofTregcells,Foxp3mRNAlevel,andIL-10expressionlevel.

Th17cellsareasubsetofCD4+Tcellsthatsecretepro-inflammatorycytokines,includingIL-17,andplayakeyroleinthepathogenesisofasthma.Tregcells,ontheotherhand,areasubsetofCD4+Tcellsthatsuppresstheimmuneresponseandmaintainimmunehomeostasis.ThebalancebetweenTh17cellsandTregcellsiscrucialfortheregulationoftheimmuneresponseinasthma.

Inaddition,HuangqiJiaqinalsoexhibitedanti-oxidativestresseffectsbyincreasingtheactivityofSODandlevelsofGSHandT-AOCinasthmaticmouselungtissue.Oxidativestressisakeyfactorinthepathogenesisofasthma,whichcancauseairwayinflammationandbronchialhyperresponsiveness.

Therefore,ourfindingssuggestthatHuangqiJiaqincanregulateimmunityandrelieveoxidativestress,therebyexertingtherapeuticeffectsonasthma.TheseresultsprovideascientificbasisfortheclinicalapplicationofHuangqiJiaqininthetreatmentofasthma.FurtherstudiesareneededtoelucidatethedetailedmechanismsofactionofHuangqiJiaqininasthmaandtoexploreitspotentialclinicalapplicationsInadditiontothemechanismsdiscussedabove,otherstudieshavealsosuggestedpotentialtherapeuticeffectsofHuangqiJiaqinonasthmathroughvariouspathways.

OnestudyfoundthatHuangqiJiaqincaninhibittheexpressionofmatrixmetalloproteinase-9(MMP-9)andreduceairwayremodelinginamousemodelofasthma(Liuetal.,2018).MMP-9isanenzymethatplaysacrucialroleinthedegradationofextracellularmatrix,anditsoverexpressionhasbeenassociatedwithairwayremodelinginasthma(Chettaetal.,2005).Therefore,theinhibitionofMMP-9byHuangqiJiaqincouldpotentiallypreventthedevelopmentofairwayremodelinginasthmaticpatients.

AnotherstudydemonstratedthatHuangqiJiaqincanregulatethelevelsofTh17cellsandIL-17inamousemodelofasthma(Zhangetal.,2015).Th17cellsareatypeofThelpercellthatproducesIL-17,apro-inflammatorycytokinethathasbeenimplicatedinthepathogenesisofasthma(KollsandLinden,2004).Therefore,theregulationofTh17cellsandIL-17byHuangqiJiaqincouldpotentiallyreduceairwayinflammationinasthmaticpatients.

Furthermore,HuangqiJiaqinhasbeenshowntoincreasethelevelsofprostacyclin(PGI2)andreducethelevelsofleukotrieneB4(LTB4)inaratmodelofasthma(Zhangetal.,2012).PGI2isavasodilatorandinhibitorofplateletaggregation,whileLTB4isapotentchemoattractantforneutrophilsandeosinophils(RouzerandKuhn,2002).TheimbalancebetweenPGI2andLTB4hasbeensuggestedtocontributetothepathogenesisofasthma(Gauvreauetal.,2010).Therefore,themodulationofPGI2andLTB4byHuangqiJiaqincouldpotentiallyimproveasthmasymptoms.

Inconclusion,HuangqiJiaqinhasbeenshowntoregulateimmunity,reduceoxidativestress,inhibitairwayinflammation,andpotentiallypreventairwayremodelinginvariousanimalstudiesofasthma.ThesefindingsprovideascientificbasisfortheclinicalapplicationofHuangqiJiaqininthetreatmentofasthma,althoughfurtherstudiesareneededtoelucidateitsdetailedmechanismsofactionandexploreitsclinicalapplicationsAdditionally,studieshavealsoshownthatHuangqiJiaqincanimprovelungfunctionandreducetheuseofrescueinhalersinpatientswithasthma.Inarandomizedcontrolledtrial,patientswithmildtomoderateasthmaweregiveneitherHuangqiJiaqinoraplacebofor12weeks.ThegroupthatreceivedHuangqiJiaqinshowedsignificantimprovementsinlungfunction,asmeasuredbyFEV1andFVC,andareductioninrescueinhalerusecomparedtotheplacebogroup.

Furthermore,HuangqiJiaqinhasalsobeenfoundtohaveasynergisticeffectwithconventionalasthmatreatments,suchasinhaledcorticosteroidsandbronchodilators.Inastudyofpatientswithsevereasthma,theadditionofHuangqiJiaqintotheirstandardtreatmentregimenresultedinasignificantdecreaseinsymptomsandimprovedlungfunction.

Overall,HuangqiJiaqinappearstobeapromisingnaturalremedyforthetreatmentofasthma.Itsabilitytoregulateimmunity,reduceoxidat