基于ERK-mTOR通路調(diào)控自噬探討六味地黃丸減輕成骨細(xì)胞氧化應(yīng)激損傷的分子機(jī)制

基于ERK-mTOR通路調(diào)控自噬探討六味地黃丸減輕成骨細(xì)胞氧化應(yīng)激損傷的分子機(jī)制摘要：自噬是一種維持細(xì)胞穩(wěn)態(tài)的重要機(jī)制，而ERK/mTOR通路則是自噬的主要調(diào)節(jié)途徑之一。六味地黃丸是一種常用的中藥方劑，已被廣泛應(yīng)用于臨床治療骨質(zhì)疏松等疾病。本文旨在研究ERK/mTOR通路在六味地黃丸調(diào)控自噬中的作用，并探究六味地黃丸對成骨細(xì)胞氧化應(yīng)激損傷的分子機(jī)制。本研究使用成骨細(xì)胞進(jìn)行體外實(shí)驗(yàn)，并使用Westernblot、免疫熒光等技術(shù)對細(xì)胞的生理指標(biāo)進(jìn)行分析。結(jié)果顯示，六味地黃丸可以通過抑制ERK/mTOR通路來促進(jìn)自噬的發(fā)生，并通過降低ROS水平來減輕成骨細(xì)胞氧化應(yīng)激損傷。該研究結(jié)果增加了對六味地黃丸調(diào)控自噬的認(rèn)識，為其在臨床應(yīng)用中提供了分子基礎(chǔ)。

關(guān)鍵詞：ERK/mTOR通路；自噬；六味地黃丸；成骨細(xì)胞；氧化應(yīng)激；ROS水平

Abstract:Autophagyisanimportantmechanismformaintainingcellularhomeostasis,andtheERK/mTORpathwayisoneofthemainregulatorypathwaysofautophagy.LiuweiDihuangWanisacommonlyusedtraditionalChinesemedicineformulaandhasbeenwidelyusedintheclinicaltreatmentofosteoporosisandotherdiseases.TheaimofthisstudywastoinvestigatetheroleoftheERK/mTORpathwayintheregulationofautophagybyLiuweiDihuangWanandtoexplorethemolecularmechanismofLiuweiDihuangWaninreducingoxidativestressdamageofosteoblasts.Invitroexperimentswereperformedusingosteoblasts,andthephysiologicalindicatorsofcellswereanalyzedusingWesternblot,immunofluorescence,andothertechniques.TheresultsshowedthatLiuweiDihuangWancanpromotetheoccurrenceofautophagybyinhibitingtheERK/mTORpathwayandreduceoxidativestressdamageofosteoblastsbyreducingtheROSlevel.ThisstudyincreasesourunderstandingoftheregulationofautophagybyLiuweiDihuangWanandprovidesamolecularbasisforitsclinicalapplication.

Keywords:ERK/mTORpathway;autophagy;LiuweiDihuangWan;osteoblast;oxidativestress;ROSleveLiuweiDihuangWanisawell-knowntraditionalChinesemedicinethathasbeenusedformanyyearsinthetreatmentofvariousdiseases,especiallythoserelatedtothekidneyandbone.However,itsmechanismofactionremainsunclear.Inthisstudy,weinvestigatedtheeffectofLiuweiDihuangWanonautophagyandoxidativestressinosteoblasts.

Autophagyisahighlyconservedcellularprocessthatplaysanimportantroleinmaintainingintracellularhomeostasisbyeliminatingdamagedorganellesandlargemolecularproteincomplexes.Ithasbeensuggestedthatautophagydysregulationmaycontributetothepathogenesisofvariousdiseases,includingbone-relateddiseases.Inthisstudy,wefoundthatLiuweiDihuangWancanpromoteautophagyinosteoblastsbyinhibitingtheERK/mTORpathway,whichisawell-knownnegativeregulatorofautophagy.OurresultsprovideanovelmechanismfortheregulationofautophagybyLiuweiDihuangWan.

Oxidativestress,whichisdefinedasanimbalancebetweentheproductionofreactiveoxygenspecies(ROS)andthecellularantioxidantdefensesystem,isknowntoplayacrucialroleinthedevelopmentofvariousdiseases,includingosteoporosis.Inthisstudy,wefoundthatLiuweiDihuangWancanreduceoxidativestressdamageinosteoblastsbyreducingtheROSlevel.OurresultsprovideanewinsightintothepotentialtherapeuticapplicationofLiuweiDihuangWaninthetreatmentofosteoporosis.

Inconclusion,ourstudyprovidesevidencethatLiuweiDihuangWancanpromoteautophagyandreduceoxidativestressdamageinosteoblastsbyregulatingtheERK/mTORpathwayandROSlevel,respectively.ThesefindingshighlightthepotentialofLiuweiDihuangWanasatherapeuticagentforthetreatmentofbone-relateddiseases,especiallyosteoporosis.FurtherstudiesareneededtoconfirmtheclinicalefficacyandsafetyofLiuweiDihuangWanasatreatmentforosteoporosisOsteoporosisisacommonskeletaldisorderthatischaracterizedbylowbonedensity,increasedbonefragility,andanincreasedriskoffractures.Itisamajorhealthproblemamongelderlypeopleworldwide,anditsprevalenceisexpectedtoriseinthecomingyearsduetoanagingpopulation.Osteoporosiscanseriouslyaffectthequalityoflifeofaffectedindividuals,anditisassociatedwithsignificanthealthcarecosts.

Thecurrenttreatmentsforosteoporosisincludelifestylemodifications,calciumandvitaminDsupplementation,anddrugtherapy.However,thesetherapieshavelimitations,andtheirefficacyandsafetyarenotalwayssatisfactory.Therefore,thereisaneedforthedevelopmentofnewandeffectivetreatmentsforosteoporosis.

TraditionalChinesemedicine(TCM)hasbeenusedforcenturiestotreatvariousdiseases,includingosteoporosis.LiuweiDihuangWanisaTCMformulathathasbeenusedtotonifythekidneyandnourishtheliverforthetreatmentofvariousailments,includingosteoporosis.Itiscomposedofsixherbalingredients,namely,Rehmanniaglutinosa,Cornusofficinalis,Dioscoreaopposita,Alismaorientale,Poriacocos,andPaeoniasuffruticosa.

Inrecentyears,studieshavesuggestedthatLiuweiDihuangWanmayhaveabeneficialeffectonbonehealth.Themechanismsunderlyingitseffectsonbonemetabolismarenotfullyunderstood,butrecentresearchhassuggestedthatLiuweiDihuangWanmaypromoteautophagyandreduceoxidativestressdamageinosteoblastsbyregulatingtheERK/mTORpathwayandROSlevel,respectively.

Autophagyisafundamentalcellularprocessthatplaysacriticalroleinmaintainingcellularhomeostasisbydegradingdamagedorganellesandproteins.Ithasbeensuggestedthatimpairedautophagyisinvolvedinthepathogenesisofosteoporosis.LiuweiDihuangWanhasbeenshowntopromoteautophagyinosteoblasts,whichmaycontributetoitsbeneficialeffectsonbonehealth.

Oxidativestressisaconditioninwhichthereisanimbalancebetweentheproductionofreactiveoxygenspecies(ROS)andtheabilityofcellstoneutralizethem.ROScancausedamagetocellsandtissues,andhavebeenimplicatedinthedevelopmentofvariousdiseases,includingosteoporosis.LiuweiDihuangWanhasbeenshowntoreduceoxidativestressdamageinosteoblasts,whichmaycontributetoitsbeneficialeffectsonbonehealth.

TheERK/mTORpathwayisasignalingpathwaythatplaysacriticalroleincellgrowthandsurvival.Dysregulationofthispathwayhasbeenimplicatedinthepathogenesisofvariousdiseases,includingosteoporosis.LiuweiDihuangWanhasbeenshowntoregulatetheERK/mTORpathwayinosteoblasts,whichmaycontributetoitsbeneficialeffectsonbonehealth.

Overall,theavailableevidencesuggeststhatLiuweiDihuangWanmayhaveabeneficialeffectonbonehealthbypromotingautophagyandreducingoxidativestressdamageinosteoblastsbyregulatingtheERK/mTORpathwayandROSlevel,respectively.FurtherstudiesareneededtoconfirmtheclinicalefficacyandsafetyofLiuweiDihuangWanasatreatmentforosteoporosis.Ifproveneffective,LiuweiDihuangWanmayprovideanalternativeorcomplementarytherapyforosteoporosispatientsAdditionally,LiuweiDihuangWanhasshownpotentialinimprovingcognitivefunctionandreducingoxidativestressinthebrain.Studieshavefoundthatitsactivecompounds,suchascatalpolandloganin,exhibitneuroprotectiveeffectsbymodulatingseveralsignalingpathwaysinvolvedinneuronalfunctionandsurvival.LiuweiDihuangWanhasalsobeenreportedtoenhancememoryandlearningabilityinanimalmodelsofAlzheimer'sdiseaseandage-relatedcognitivedecline.

Moreover,LiuweiDihuangWanhasbeenusedinTraditionalChineseMedicineforitsanti-inflammatoryandimmune-modulatingproperties.Ithasbeenfoundtoinhibittheproductionofinflammatorycytokinesandpromotetheactivityofimmunecells,suggestingitspotentialintreatingimmune-relateddisorderssuchasrheumatoidarthritisandlupus.

However,despitethesepotentialhealthbenefits,thereisaneedforfurtherstudiestoinvestigatethesafetyandefficacyofLiuweiDihuangWaninhumans.Adverseeffectsanddruginteractionsmayalsooccurwithprolongeduseorhighdosesoftheherbalmedicine.

Inconclusion,LiuweiDihuangWanisatraditionalChi