基于PI3K-Akt信號通路探討百會穴按摩對MCAO大鼠認知功能影響的作用機制

基于PI3K-Akt信號通路探討百會穴按摩對MCAO大鼠認知功能影響的作用機制摘要：目的：探討基于PI3K/Akt信號通路的百會穴按摩對中大腦動脈阻塞（MCAO）大鼠認知功能的影響及其作用機制。方法：選取60只健康雄性SD大鼠，隨機分為正常組、MCAO組、百會穴按摩組、LY294002組和Massage+LY294002組，其中MCAO組、百會穴按摩組、Massage+LY294002組采用MCAO大鼠模型，正常組不做任何處理；百會穴按摩組和Massage+LY294002組于MCAO后于百會穴進行按摩（每天30min，連續(xù)7天），LY294002組注射LY294002；Morris水迷宮測試評估大鼠認知功能，RT-qPCR技術檢測PI3K和Akt基因表達，Westernblot檢測相關蛋白的表達水平。結果：MCAO組大鼠在Morris水迷宮測試中表現(xiàn)出明顯的認知功能障礙，百會穴按摩和LY294002可顯著改善其認知功能（P<0.05）；PI3K和Akt基因表達和相關蛋白的表達水平在各組之間存在差異（P<0.05），百會穴按摩可通過激活PI3K/Akt信號通路來提高認知功能。結論：百會穴按摩可通過激活PI3K/Akt信號通路來改善MCAO大鼠的認知功能。關鍵詞：中大腦動脈阻塞；百會穴按摩；PI3K/Akt信號通路；認知功能。

Abstract:Objective:ToexploretheeffectandmechanismofBaihuiacupointmassagebasedonPI3K/Aktsignalingpathwayoncognitivefunctionofmiddlecerebralarteryocclusion(MCAO)rats.Methods:SixtyhealthymaleSDratswererandomlydividedintoanormalgroup,MCAOgroup,Baihuiacupointmassagegroup,LY294002group,andMassage+LY294002group.MCAO,Baihuiacupointmassage,andMassage+LY294002groupswereinducedwithMCAOratmodelandmassagefor30minperdayfor7consecutivedaysafterMCAO.LY294002groupwasinjectedwithLY294002.TheMorriswatermazetestwasusedtoevaluatethecognitivefunctionofrats.RT-qPCRandWesternblotwereusedtodetecttheexpressionofPI3KandAktgenesandrelevantproteins.Results:MCAOgroupratsexhibitedobviouscognitivedysfunctionintheMorriswatermazetest,andBaihuiacupointmassageandLY294002significantlyimprovedthecognitivefunction(P<0.05).ThereweredifferencesinPI3KandAktgeneexpressionandrelevantproteinexpressionlevelsamonggroups(P<0.05),andBaihuiacupointmassagecanimprovecognitivefunctionbyactivatingthePI3K/Aktsignalingpathway.Conclusion:BaihuiacupointmassagecanimprovecognitivefunctionofMCAOratsbyactivatingthePI3K/Aktsignalingpathway.Keywords:middlecerebralarteryocclusion;Baihuiacupointmassage;PI3K/Aktsignalingpathway;cognitivefunctionInconclusion,thepresentstudydemonstratesthatBaihuiacupointmassagecanimprovecognitivefunctioninMCAOratsbyactivatingthePI3K/Aktsignalingpathway.TheresultsindicatethatthetherapeuticeffectofBaihuiacupointmassageoncognitiveimpairmentisassociatedwiththemodulationofPI3K/Aktsignalingpathwayactivity.ThesefindingssuggestthatBaihuiacupointmassagecouldbeapotentialtherapeuticstrategyforthetreatmentofcognitivedysfunctioninstrokepatients.

Previousstudieshaveshownthatotheracupuncturetechniques,suchaselectroacupunctureandmanualacupuncture,canalsoimprovecognitivefunctioninanimalmodelsofstroke(Liuetal.,2015;Wangetal.,2015b).Thesefindingssuggestthatacupuncturetherapiesmaybeusefulfortreatingcognitiveimpairmentinstrokepatients.However,furtherstudiesarerequiredtodeterminetheoptimalacupuncturetechniqueandtreatmentdurationforstrokepatients.

Thepresentstudyhassomelimitations.Firstly,themechanismunderlyingtheeffectofBaihuiacupointmassageonthePI3K/Aktsignalingpathwaywasnotfullyelucidated.FuturestudiesshouldinvestigatethesignalingpathwaysdownstreamofPI3K/AktthatmaybeimplicatedintheeffectofBaihuiacupointmassageoncognitivefunction.Secondly,thesamplesizewasrelativelysmall,whichmaylimitthegeneralizabilityofthefindings.Finally,thestudyonlyexaminedtheimmediateeffectsofBaihuiacupointmassageoncognitivefunction,andlong-termeffectswerenotevaluated.FurtherstudiesarerequiredtodeterminethedurationofthetherapeuticeffectofBaihuiacupointmassageoncognitivefunction.

Despitetheselimitations,thepresentstudyprovidesevidencethatBaihuiacupointmassagecanimprovecognitivefunctioninMCAOratsbyactivatingthePI3K/Aktsignalingpathway.Thefindingssuggestthatacupuncturetherapiesmaybeausefulstrategyfortreatingcognitivedysfunctioninstrokepatients.FurtherresearchisneededtovalidatethesefindingsandtoexploretheoptimaltreatmentregimensInadditiontotheinvestigationofthetherapeuticeffectofBaihuiacupointmassageoncognitivefunction,thereareotheracupuncturetherapiesthathavebeenexploredforstroke-relatedcognitivedysfunction.Forexample,apreviousstudyhasreportedthatelectroacupuncture(EA)attheBaihuiandDazhuiacupointscanimprovecognitiveimpairmentinstrokepatientsbyupregulatingBDNFexpression(Wangetal.,2012).Anotherstudyfoundthatscalpacupuncturecombinedwithcognitivetrainingcanimprovecognitivefunctionanddailylivingactivitiesinpost-strokepatientswithvasculardementia(Lietal.,2016).Moreover,acupuncturetherapieshavebeenshowntoberelativelysafeandwell-toleratedinstrokepatients,withfewadverseeffectsreported(Liuetal.,2018).

Despitethepromisingresults,thereareseveralchallengesthatneedtobeaddressedinfutureresearch.First,themechanismsunderlyingthetherapeuticeffectsofacupuncturetherapiesoncognitivefunctionafterstrokeremainpoorlyunderstood.Second,thereisalackofstandardizedtreatmentprotocolsandwell-designedrandomizedcontrolledtrialsthatareneededtoestablishtheoptimalacupuncturetreatmentregimensforstroke-relatedcognitivedysfunction.Third,thegeneralizabilityofthesefindingsmaybelimitedbytheheterogeneityofstrokepatientsandthediversityofacupuncturetechniquesusedindifferentstudies.Therefore,asystematicandcomprehensiveapproachthatintegratesclinical,biological,andneuroimagingmeasuresisneededtoelucidatethemechanismsofacupuncturetherapiesandtooptimizetheirclinicalefficacyforstroke-relatedcognitivedysfunction.

Insummary,thepresentstudysupportstheuseofBaihuiacupointmassageasaneffectiveandsafetreatmentstrategyforcognitivedysfunctionafterstroke,andhighlightsthepotentialroleofthePI3K/Aktsignalingpathwayinmediatingthetherapeuticeffectsofacupuncturetherapies.Thesefindingshaveimportantimplicationsforthedevelopmentofalternativeandcomplementarytherapiesforstrokepatients,andpavethewayforfurtherresearchinthisfieldCognitivedysfunctionafterstrokeisacommonanddisablingconditionthatcangreatlyimpactapatient'squalityoflife.Whilestandardmedicaltreatments,suchasmedicationsandrehabilitation,mayhelpimprovecognitivefunctiontosomeextent,theyoftencomewithlimitationsandsideeffects.Therefore,exploringalternativeandcomplementarytherapies,suchasacupuncture,couldoffernewinsightsintothemanagementofpost-strokecognitivedysfunction.

AcupunctureisatraditionalChinesetherapythatinvolvestheinsertionofthinneedlesintospecificpointsonthebodyknownasacupoints.AccordingtotraditionalChinesemedicinetheory,acupunctureworksbyregulatingtheflowofqi,orvitalenergy,throughthebody'smeridiansystem,whichinturncanhelprestorebalanceandimprovehealth.Inrecentyears,acupuncturehasgainedincreasingattentionasapotentialtreatmentforcognitivedysfunctionafterstroke,withgrowingempiricalevidencesupportingitstherapeuticeffects.

OneofthemostcommonlytargetedacupointsforcognitivedysfunctionafterstrokeisBaihui,whichislocatedonthetopofthehead.Baihuiacupointmassageisasimpleandnon-invasiveacupuncturetechniquethatinvolvesgentlepressureorcircularrubbingoftheacupointusingthefingersorasmallmassagetool.Thistechniquehasbeenshowntoimprovecognitivefunctionandreducedepressioninstrokepatients,potentiallybyenhancingbloodflowandneurotransmitteractivityinthebrain.

InadditiontoitslocaleffectsontheBaihuiacupoint,acupuncturemayexertbroadersystemiceffectsbymodulatingvarioussignalingpathwaysinthebody.OnesuchpathwaythathasbeenimplicatedinthetherapeuticeffectsofacupunctureisthePI3K/Aktpathway.Thispathwayisinvolvedinregulatingcellsurvivalandgrowth,andhasbeenshowntobeactivatedbyacupunctureinanimalandhumanstudies.Byactivatingthispathway,acupuncturemaypromotecellularandsynapticplasticityinthebrain,whichcanhelpimprovecognitivefunction.

Overall,theuseofacupuncture,particularlyBaihuiacupointmassage,appearstobeapromisingapproachforthemanagementofcognitivedysfunctionafterstroke.Whilefurtherresearchisneededtofullyelucidatetheunderlyingmech