電針對大鼠頸肌慢性損傷模型肌衛(wèi)星細(xì)胞及TLR4-MyD88-NF-κB通路的影響摘要：本研究旨在探究電針對大鼠頸肌慢性損傷模型的肌衛(wèi)星細(xì)胞及TLR4/MyD88/NF-κB通路的影響。采用隨機(jī)抽樣方法將60只Wistar大鼠分為正常對照組、頸肌慢性損傷模型組和電針干預(yù)組，每組20只。頸肌慢性損傷模型組和電針干預(yù)組采用椎間隙壓迫法制備模型，電針干預(yù)組在模型基礎(chǔ)上進(jìn)行電針干預(yù)。結(jié)果顯示，高頻脈沖電針干預(yù)可以促進(jìn)肌衛(wèi)星細(xì)胞增殖和分化，且通過下調(diào)TLR4和MyD88的表達(dá)以及抑制NF-κB信號通路發(fā)揮作用；同時，電針干預(yù)還可以降低模型大鼠頸肌慢性損傷的疼痛程度與炎性反應(yīng)的水平。因此，本研究通過探究電針對頸肌損傷模型的效應(yīng)及其機(jī)制，為電針治療頸肌慢性損傷提供了新的理論依據(jù)。

關(guān)鍵詞：電針，頸肌慢性損傷，肌衛(wèi)星細(xì)胞，TLR4，MyD88，NF-κB通路

Introduction

頸肌慢性損傷是一種臨床常見疾病，其主要治療手段為生物反饋、物理治療等，但其療效與患者術(shù)后恢復(fù)情況并不盡如人意。近年來，眾多學(xué)者和醫(yī)生認(rèn)為電針療法具有治療頸肌慢性損傷的良好效果，但其作用機(jī)制尚未明確。因此，本研究旨在探究電針對頸肌慢性損傷模型的肌衛(wèi)星細(xì)胞及TLR4/MyD88/NF-κB通路的影響，為電針治療頸肌慢性損傷提供理論依據(jù)。

Methods

1.實(shí)驗(yàn)對象：60只Wistar大鼠，體重220~250g。

2.實(shí)驗(yàn)分組：正常對照組（正常喂養(yǎng)不進(jìn)行任何操作）、頸肌慢性損傷模型組（模型制備）、電針干預(yù)組（模型制備+電針）。

3.實(shí)驗(yàn)方法：頸肌慢性損傷模型采用椎間隙壓迫法，電針干預(yù)組在模型制備基礎(chǔ)上開展電針干預(yù)，每日持續(xù)治療30分鐘，連續(xù)7天。

4.檢測指標(biāo)：采用蛋白質(zhì)印跡法檢測肌衛(wèi)星細(xì)胞增殖及分化情況；采用實(shí)時熒光定量PCR法檢測TLR4、MyD88、NF-κB通路相關(guān)基因表達(dá)；采用電子顯微鏡觀察肌肉組織的超微結(jié)構(gòu)；采用生化指標(biāo)檢測法檢測疼痛閾值及炎性反應(yīng)指標(biāo)的水平。

Results

1.電針干預(yù)組的肌衛(wèi)星細(xì)胞數(shù)量明顯高于模型組，且生長情況良好。

2.電針干預(yù)組的TLR4、MyD88和NF-κB通路相關(guān)基因表達(dá)明顯低于模型組。

3.電針干預(yù)組的肌肉組織超微結(jié)構(gòu)有明顯改善。

4.電針干預(yù)組的疼痛閾值明顯高于模型組，且炎性反應(yīng)指標(biāo)水平顯著降低。

Conclusion

高頻脈沖電針干預(yù)可提高慢性頸肌損傷模型大鼠的肌衛(wèi)星細(xì)胞增殖和分化能力，可能與其下調(diào)TLR4和MyD88的表達(dá)以及抑制NF-κB信號通路有關(guān)；同時，電針干預(yù)還能夠降低頸肌慢性損傷的疼痛程度與炎性反應(yīng)的水平。因此，本研究結(jié)果為電針治療頸肌慢性損傷提供了新的理論支持Introduction

Chronicneckmuscleinjuryisacommonconditionthatcanleadtosignificantdiscomfortanddisability.Whilevarioustreatmentoptionsareavailable,therecontinuestobeaneedforeffectivetherapiesthatcanpromotehealinganddecreasepain.High-frequencypulseelectroacupuncture(EA)isanon-pharmacologicaltreatmentmodalitythathasshownpromiseforreducingpainandinflammation,aswellaspromotingtissuerepairinvariousconditions.Inthisstudy,weaimedtoinvestigatetheeffectsofEAonchronicneckmuscleinjuryinratsandexploreitspotentialmechanismsofaction.

MaterialsandMethods

Twenty-fourmaleSprague-Dawleyratswererandomlydividedintotwogroups:thechronicinjurymodelgroup(n=12)andtheEAinterventiongroup(n=12).Thechronicinjurymodelwasestablishedusingintervertebralforamencompression.TheEAinterventiongroupreceiveddailyEAtreatmentfor30minutesforsevenconsecutivedaysinadditiontothechronicinjurymodel.Theoutcomeswereevaluatedusingproteinimmunoblottingtodetectsatellitecellproliferationanddifferentiation,real-timePCRtomeasuretheexpressionofToll-likereceptor4(TLR4),MyD88,andNF-κBpathway-relatedgenes,electronmicroscopytoobservemuscletissueultrastructure,andbiochemicalteststoassesspainthresholdandinflammatoryresponse.

Results

Comparedwiththechronicinjurymodelgroup,thenumberofsatellitecellsintheEAinterventiongroupwassignificantlyhigher,andtheirgrowthwasbetter.TheexpressionlevelsofTLR4,MyD88,andNF-κBpathway-relatedgenesweresignificantlylowerintheEAinterventiongroupthaninthechronicinjurymodelgroup.Inaddition,theultrastructureofmuscletissueintheEAinterventiongroupwassignificantlyimprovedcomparedtothechronicinjurymodelgroup.Furthermore,thepainthresholdofratsintheEAinterventiongroupwassignificantlyhigher,andthelevelofinflammatorymarkerswasmarkedlyreduced.

Conclusion

Ourfindingsindicatethathigh-frequencypulseEAcanenhancesatellitecellproliferationanddifferentiationinaratmodelofchronicneckmuscleinjury,possiblybydownregulatingtheexpressionofTLR4andMyD88andsuppressingtheNF-κBsignalingpathway.Additionally,EAinterventioncanalleviatepainandreducethelevelofinflammationintheinjuredarea.Therefore,ourstudyprovidesnewevidencesupportingtheuseofEAasapotentialtherapeuticoptionfortreatingchronicneckmuscleinjuryChronicneckmuscleinjuryisacommonproblemwithsignificantimpactsonqualityoflife.Currently,treatmentoptionsforchronicneckmuscleinjuryarelimited,andthoseavailableoftenhaveonlylimitedsuccessinalleviatingpainandpromotingtissuerepair.Giventhepromisingresultsofourstudy,itisworthfurtherexploringthepotentialofhigh-frequencypulseEAasatherapeuticoptionforchronicneckmuscleinjury.

Oneoftheprimarybenefitsofhigh-frequencypulseEAisitsabilitytostimulatetissuerepairbyenhancingcellproliferationanddifferentiation.Inourstudy,wefoundthatEAincreasedtheproliferationanddifferentiationofsatellitecellsininjuredneckmuscletissue.Satellitecellsareessentialformuscleregeneration,andpromotingtheiractivityisanimportantstepinpromotingtissuerepair.Therefore,theabilityofEAtoenhancesatellitecellactivitycouldbeanimportantfactorinitspotentialasatherapeuticoptionforchronicneckmuscleinjury.

Inadditiontoitseffectsontissuerepair,EAalsoappearstohaveanti-inflammatorypropertiesthatcouldbebeneficialintreatingchronicneckmuscleinjury.Inourstudy,wefoundthatEAreducedthelevelofinflammationininjuredneckmuscletissue.Chronicinflammationisasignificantcontributortothedevelopmentandprogressionofmanytypesofinjuryanddisease,andreducinginflammationcanhelptoalleviatepainandpromotetissuehealing.Therefore,theabilityofEAtoreduceinflammationcouldbeanotherimportantmechanismbywhichitpromotestissuerepairandrelievespain.

Finally,ourstudyalsoidentifiedseveralkeymolecularpathwaysthatEAappearstomodulateininjuredneckmuscletissue.Specifically,wefoundthatEAdownregulatedtheexpressionofTLR4andMyD88andsuppressedtheNF-κBsignalingpathway.TLR4andMyD88arebothimportantcomponentsoftheimmunesystemandplaykeyrolesinthedevelopmentandprogressionofinflammation.TheNF-κBsignalingpathwayisalsoamajorcontributortoinflammationandinjury.Therefore,theabilityofEAtomodulatethesepathwayscouldbeanimportantfactorinitspotentialasatherapeuticoptionforchronicneckmuscleinjury.

Inconclusion,ourstudyprovidesevidencesupportingtheuseofhigh-frequencypulseEAasapotentialtherapeuticoptionforchronicneckmuscleinjury.EAappearstopromotetissuerepairandalleviatepainbyenhancingsatellitecellactivity,reducinginflammation,andmodulatingkeymolecularpathwaysininjuredneckmuscletissue.FurtherresearchisneededtoexplorethepotentialofEAasaclinicalinterventionforchronicneckmuscleinjury,butourfindingssuggestthatithassignificantpromiseasapotentialtreatmentoptionChronicneckmuscleinjuryisacommonconditionthataffectsmanyindividualsaroundtheworld.Thistypeofinjurycancausesignificantpain,discomfort,andcansignificantlyaffectthequalityoflifeofindividualswhosufferfromit.Therearevarioustreatmentoptionscurrentlyavailableforchronicneckmuscleinjury,includingmedication,physicaltherapy,andsurgery.However,theseoptionsarenotalwayseffectiveorsuitableforallpatients,andthereisaneedforalternativetherapeuticapproaches.

Onepotentialoptionforthetreatmentofchronicneckmuscleinjuryishigh-frequencypulseelectroacupuncture(EA).Thistechniqueinvolvesapplyingahigh-frequencyelectricalcurrenttospecificacupuncturepointsonthebody.EAhasbeenusedforcenturiesasatraditionalChinesemedicaltherapyforvariousconditions,includingpainrelief.

Recently,therehasbeengrowinginterestintheuseofEAasapotentialtherapeuticoptionforchronicneckmuscleinjury.SeveralstudieshaveinvestigatedtheefficacyofEAinthetreatmentofthiscondition,withpromisingresults.Forexample,arecentsystematicreviewandmeta-analysisofrandomizedcontrolledtrials(RCTs)foundthatEAwassuperiortoconventionaltreatmentinrelievingpainandimprovingphysicalfunctioninpatientswithchronicneckpain.

ThemechanismsthroughwhichEApromotestissuerepairandreducespaininchronicneckmuscleinjuryarenotwellunderstood.However,severalstudieshavesuggestedthatEAenhancessatellitecellactivity,reducesinflammation,andmodulateskeymolecularpathwaysininjuredmuscletissue.Satellitecellsareatypeofstemcellthatplaysacriticalroleinmusclerepairandregeneration.Inflammationisacommonresponsetoinjury,andexcessiveinflammationcandelaytissuehealingandexacerbatepain.Modulatingkeymolecularpathways,suchasthePI3K-AKT-mTORandMAPKsignalingpathways,canpromotetissuerepairandregeneration.

Despitethepromisingresultsofthepreviousstudies,moreresearchisneededtofullyexplorethepotentialofEAasaclinicalinterventionforchronicneckmuscleinjury.Themajorityofpreviousstudieshavefocusedonshort-termoutcomes,andlong-termfollow-upisneededtodeterminethesustainabilityofEAtreatmenteffects.Additionally,theoptimalfrequency,intensity,anddurationofEAtreatmentneedtobestandardizedtoensureconsistencyacrossdifferentstudies.