电针对大鼠颈肌慢性损伤模型肌卫星细胞及TLR4-MyD88-NF-κB通路的影响

电针对大鼠颈肌慢性损伤模型肌卫星细胞及TLR4-MyD88-NF-κB通路的影响摘要：本研究旨在探究电针对大鼠颈肌慢性损伤模型的肌卫星细胞及TLR4/MyD88/NF-κB通路的影响。采用随机抽样方法将60只Wistar大鼠分为正常对照组、颈肌慢性损伤模型组和电针干预组，每组20只。颈肌慢性损伤模型组和电针干预组采用椎间隙压迫法制备模型，电针干预组在模型基础上进行电针干预。结果显示，高频脉冲电针干预可以促进肌卫星细胞增殖和分化，且通过下调TLR4和MyD88的表达以及抑制NF-κB信号通路发挥作用；同时，电针干预还可以降低模型大鼠颈肌慢性损伤的疼痛程度与炎性反应的水平。因此，本研究通过探究电针对颈肌损伤模型的效应及其机制，为电针治疗颈肌慢性损伤提供了新的理论依据。

关键词：电针，颈肌慢性损伤，肌卫星细胞，TLR4，MyD88，NF-κB通路

Introduction

颈肌慢性损伤是一种临床常见疾病，其主要治疗手段为生物反馈、物理治疗等，但其疗效与患者术后恢复情况并不尽如人意。近年来，众多学者和医生认为电针疗法具有治疗颈肌慢性损伤的良好效果，但其作用机制尚未明确。因此，本研究旨在探究电针对颈肌慢性损伤模型的肌卫星细胞及TLR4/MyD88/NF-κB通路的影响，为电针治疗颈肌慢性损伤提供理论依据。

Methods

1.实验对象：60只Wistar大鼠，体重220~250g。

2.实验分组：正常对照组（正常喂养不进行任何操作）、颈肌慢性损伤模型组（模型制备）、电针干预组（模型制备+电针）。

3.实验方法：颈肌慢性损伤模型采用椎间隙压迫法，电针干预组在模型制备基础上开展电针干预，每日持续治疗30分钟，连续7天。

4.检测指标：采用蛋白质印迹法检测肌卫星细胞增殖及分化情况；采用实时荧光定量PCR法检测TLR4、MyD88、NF-κB通路相关基因表达；采用电子显微镜观察肌肉组织的超微结构；采用生化指标检测法检测疼痛阈值及炎性反应指标的水平。

Results

1.电针干预组的肌卫星细胞数量明显高于模型组，且生长情况良好。

2.电针干预组的TLR4、MyD88和NF-κB通路相关基因表达明显低于模型组。

3.电针干预组的肌肉组织超微结构有明显改善。

4.电针干预组的疼痛阈值明显高于模型组，且炎性反应指标水平显著降低。

Conclusion

高频脉冲电针干预可提高慢性颈肌损伤模型大鼠的肌卫星细胞增殖和分化能力，可能与其下调TLR4和MyD88的表达以及抑制NF-κB信号通路有关；同时，电针干预还能够降低颈肌慢性损伤的疼痛程度与炎性反应的水平。因此，本研究结果为电针治疗颈肌慢性损伤提供了新的理论支持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.