基于PI3K-Akt信號(hào)通路探討百會(huì)穴按摩對(duì)MCAO大鼠認(rèn)知功能影響的作用機(jī)制摘要：目的：探討基于PI3K/Akt信號(hào)通路的百會(huì)穴按摩對(duì)中大腦動(dòng)脈阻塞（MCAO）大鼠認(rèn)知功能的影響及其作用機(jī)制。方法：選取60只健康雄性SD大鼠，隨機(jī)分為正常組、MCAO組、百會(huì)穴按摩組、LY294002組和Massage+LY294002組，其中MCAO組、百會(huì)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(huì)穴按摩組和Massage+LY294002組于MCAO后于百會(huì)穴進(jìn)行按摩（每天30min，連續(xù)7天），LY294002組注射LY294002；Morris水迷宮測(cè)試評(píng)估大鼠認(rèn)知功能，RT-qPCR技術(shù)檢測(cè)PI3K和Akt基因表達(dá)，Westernblot檢測(cè)相關(guān)蛋白的表達(dá)水平。結(jié)果：MCAO組大鼠在Morris水迷宮測(cè)試中表現(xiàn)出明顯的認(rèn)知功能障礙，百會(huì)穴按摩和LY294002可顯著改善其認(rèn)知功能（P<0.05）；PI3K和Akt基因表達(dá)和相關(guān)蛋白的表達(dá)水平在各組之間存在差異（P<0.05），百會(huì)穴按摩可通過(guò)激活PI3K/Akt信號(hào)通路來(lái)提高認(rèn)知功能。結(jié)論：百會(huì)穴按摩可通過(guò)激活PI3K/Akt信號(hào)通路來(lái)改善MCAO大鼠的認(rèn)知功能。關(guān)鍵詞：中大腦動(dòng)脈阻塞；百會(huì)穴按摩；PI3K/Akt信號(hào)通路；認(rèn)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