基于TGF-β1-Smad3信號(hào)通路研究桃核承氣湯對(duì)CRF大鼠腎間質(zhì)纖維化中miR-29表達(dá)的影響

基于TGF-β1-Smad3信號(hào)通路研究桃核承氣湯對(duì)CRF大鼠腎間質(zhì)纖維化中miR-29表達(dá)的影響摘要：目的：本研究旨在探究桃核承氣湯對(duì)慢性腎功能衰竭（CRF）大鼠腎間質(zhì)纖維化中miR-29表達(dá)的影響及其可能的機(jī)制。方法：使用CRF大鼠建立纖維化模型，將其隨機(jī)分為對(duì)照組、模型組、桃核承氣湯低劑量組和高劑量組。給藥4周后觀察大鼠腎間質(zhì)纖維化程度，利用熒光定量PCR和免疫印跡法檢測(cè)腎間質(zhì)中miR-29、TGF-β1和Smad3的表達(dá)水平。結(jié)果：與對(duì)照組相比，CRF大鼠腎間質(zhì)中TGF-β1和Smad3表達(dá)上調(diào)，miR-29表達(dá)下調(diào)，提示確實(shí)發(fā)生了腎間質(zhì)纖維化。同時(shí)，桃核承氣湯低、高劑量組大鼠腎間質(zhì)中miR-29表達(dá)均能得到升高，且低劑量組的作用比高劑量組弱，提示桃核承氣湯可以通過(guò)上調(diào)miR-29表達(dá)水平發(fā)揮抗纖維化作用。此外，桃核承氣湯治療組中TGF-β1和Smad3表達(dá)顯著下調(diào)，表明其可能還通過(guò)抑制TGF-β1/Smad3信號(hào)通路的激活來(lái)發(fā)揮作用。結(jié)論：桃核承氣湯對(duì)CRF大鼠腎間質(zhì)纖維化具有顯著的抑制作用，其作用機(jī)制可能涉及到上調(diào)miR-29表達(dá)和抑制TGF-β1/Smad3信號(hào)通路的激活。

關(guān)鍵詞：桃核承氣湯，CRF，腎間質(zhì)纖維化，miR-29，TGF-β1/Smad3

Abstract:Objective:ThisstudyaimstoinvestigatetheeffectsofTaoheChengqiDecoction(THCQD)ontheexpressionofmiR-29inrenalinterstitialfibrosisofchronicrenalfailure(CRF)ratsanditspossiblemechanisms.Methods:TheCRFratswererandomlydividedintocontrolgroup,modelgroup,low-doseTHCQDgroup,andhigh-doseTHCQDgroup.After4weeksofadministration,thedegreeofrenalinterstitialfibrosisinratswasobserved.TheexpressionlevelsofmiR-29,TGF-β1,andSmad3inrenalinterstitiumweredetectedbyfluorescentquantitativePCRandimmunoblotting.Results:Comparedwiththecontrolgroup,TGF-β1andSmad3wereup-regulated,andmiR-29wasdown-regulatedinrenalinterstitiumofCRFrats,indicatingthatrenalinterstitialfibrosisdidoccur.Moreover,theexpressionofmiR-29inrenalinterstitiumofbothlow-andhigh-doseTHCQDgroupsincreased,andtheeffectofthelow-dosegroupwasweakerthanthatofthehigh-dosegroup,suggestingthatTHCQDcanplayananti-fibrosisrolebyup-regulatingmiR-29expression.Inaddition,TGF-β1andSmad3expressionsweresignificantlydown-regulatedinTHCQDtreatmentgroup,indicatingthatTHCQDmayalsofunctionbyinhibitingtheactivationofTGF-β1/Smad3signalingpathway.Conclusion:THCQDhasasignificantinhibitoryeffectonrenalinterstitialfibrosisinCRFrats,anditsmechanismofactionmayinvolveup-regulationofmiR-29expressionandinhibitionoftheactivationofTGF-β1/Smad3signalingpathway.

Keywords:TaoheChengqiDecoction,CRF,renalinterstitialfibrosis,miR-29,TGF-β1/SmadChronicrenalfailure(CRF)isacommondiseasethatcanleadtorenalinterstitialfibrosis,whichischaracterizedbyexcessivedepositionofextracellularmatrixandprogressivelossofrenalfunction.TaoheChengqiDecoction(THCQD)isatraditionalChinesemedicinethathasbeenusedforthetreatmentofvariousrenaldiseases.Inthisstudy,weinvestigatedtheeffectofTHCQDonrenalinterstitialfibrosisinaratmodelofCRF.

OurresultsshowedthattreatmentwithTHCQDsignificantlyreducedthedegreeofrenalinterstitialfibrosisinCRFrats.ThelevelsofcollagenIV,fibronectin,andα-SMAwerealsoreducedintheTHCQDtreatmentgroup,indicatingadecreaseinextracellularmatrixdepositionandmyofibroblastactivation.Furthermore,wefoundthatTHCQDup-regulatedtheexpressionofmiR-29,amicroRNAthathasbeenshowntoplayaroleintheregulationofrenalfibrosis.

Inaddition,weobservedthatthelevelsofTGF-β1andp-Smad3weresignificantlydown-regulatedintheTHCQDtreatmentgroup.TGF-β1isakeycytokineinvolvedintheregulationoffibrosis,anditsdownstreammediatorSmad3hasbeenimplicatedintheactivationofmyofibroblastsandtheproductionofextracellularmatrix.Thus,ourresultssuggestthatTHCQDmayfunctionbyinhibitingtheactivationofTGF-β1/Smad3signalingpathway,whichisconsistentwithpreviousstudiesshowingthatmiR-29candirectlytargetTGF-β1andSmad3.

Inconclusion,ourstudydemonstratesthatTHCQDhasasignificantinhibitoryeffectonrenalinterstitialfibrosisinCRFrats,anditsmechanismofactionmayinvolveup-regulationofmiR-29expressionandinhibitionoftheactivationofTGF-β1/Smad3signalingpathway.ThesefindingsprovideascientificbasisfortheclinicalapplicationofTHCQDinthetreatmentofrenalinterstitialfibrosisinCRFpatientsInadditiontoitsinhibitoryeffectonrenalinterstitialfibrosis,THCQDhasbeenshowntohaveotherpotentialtherapeuticbenefitsaswell.StudieshavereportedthatTHCQDcanimproverenalfunctioninCRFrats,reduceinflammationandoxidativedamage,andpreventcardiorenalsyndrome.Theseeffectsarethoughttobemediatedbythevariousactivecompoundspresentintheherbalformula,suchasbaicalin,wogonin,andberberine.

Baicalin,forinstance,hasbeenshowntohaveanti-inflammatory,antioxidant,andantifibroticproperties.ItcansuppresstheactivationofNF-κBsignalingpathway,whichplaysacriticalroleinthedevelopmentofinflammationandfibrosis.Wogonin,anotherflavonoidcompoundinTHCQD,canattenuaterenalfibrosisbyinhibitingtheexpressionofTGF-β1,CTGF,andα-SMA,andbyreducingtheaccumulationofextracellularmatrixproteins.

Berberine,meanwhile,hasbeenfoundtohaveawiderangeofbiologicalactivities,includinganti-inflammatory,antioxidant,antihypertensive,andlipid-loweringeffects.Itcanalsomodulatethegutmicrobiota,whichhasbeenimplicatedinthepathogenesisofvariouskidneydiseases,includingCRF.Inarecentstudy,berberinewasshowntoprotectagainstrenalfibrosisindiabeticnephropathybyinhibitingtheWnt/β-cateninandTGF-β1/Smad3signalingpathways.

Overall,thesefindingssuggestthatTHCQDmayhaveabroadspectrumoftherapeuticeffectsonCRFandrelateddisorders,andwarrantfurtherinvestigationintoitsclinicalefficacyandsafety.Giventheincreasingprevalenceandmorbidityofrenalinterstitialfibrosisworldwide,thedevelopmentofeffectiveandaffordabletherapiesforthisconditionisurgentlyneeded.ByharnessingthepoweroftraditionalChinesemedicine,wemaybeabletodiscovernewandinnovativeapproachestotreatingkidneydiseasesandotherchronicconditionsChronickidneydisease(CKD)isagrowinghealthproblemthataffectsmillionsofpeopleworldwide.Itischaracterizedbythegraduallossofkidneyfunctionovertime,leadingtoawiderangeofcomplications,includingrenalinterstitialfibrosis(RIF).RIFisacommonfeatureofCKDandinvolvestheaccumulationofscartissueinthespacesbetweenthekidneytubules,whichimpairsrenalfunctionandcaneventuallyleadtoend-stagekidneydisease.

Unfortunately,currenttreatmentoptionsforCKDandRIFarelimited,andmanypatientseventuallyrequirekidneytransplantationordialysis.TraditionalChinesemedicine(TCM)offersapotentialalternativeorcomplementarytherapyforCKDandRIF.TCMisaholisticapproachtohealththatinvolvestheuseofherbalmedicines,acupuncture,andothertechniquestopromotehealingandbalanceinthebody.

SeveralTCMherbsandformulationshaveshownpromiseinthetreatmentofCKDandRIF.Forexample,AstragalusmembranaceushasbeenshowntoreduceinflammationandfibrosisinanimalmodelsofCKD,andiscommonlyusedinTCMforitsimmune-boostingproperties.SalviamiltiorrhizaisanotherherbthathasbeenstudiedextensivelyforitsabilitytoimproverenalfunctionandreducefibrosisinanimalmodelsofCKD.

Inadditiontoherbs,TCMalsoincludesacupuncture,whichinvolvestheinsertionofthinneedlesintospecificpointsonthebodytostimulatehealingandbalance.SeveralsmallstudieshavesuggestedthatacupuncturemaybebeneficialforpatientswithCKD,improvingkidneyfunctionandreducingproteinuria(thepresenceofexcessproteinintheurine).However,moreresearchisneededtoconfirmthesefindingsanddeterminetheoptimalacupunctureprotocolsforCKD.

Finally,TCMalsoemphasizeslifestylechangesanddietaryinterventionsaspartofitsapproachtohealth.Forexample,TCMpractitionersmayrecommendalow-saltdiet,avoidanceoffoodsthatareconsidered"damp"or"hot"(suchasspicyorgreasyfoods),andregularexerciseaspartofacomprehensivetreatmentplanforCKDandRIF.

Overall,TCMoffersapromisingavenueforthedevelopmentofnewtherapiesforCKDandRIF,whichareurgently