基于PI3K-AKT通路探討復(fù)方斑蝥膠囊聯(lián)合阿帕替尼對(duì)骨肉瘤裸鼠模型成瘤的影響基于PI3K/AKT通路探討復(fù)方斑蝥膠囊聯(lián)合阿帕替尼對(duì)骨肉瘤裸鼠模型成瘤的影響

摘要：本研究旨在探討復(fù)方斑蝥膠囊聯(lián)合阿帕替尼對(duì)骨肉瘤的治療作用及其機(jī)制。采用H&E染色和免疫組化法觀察藥物干預(yù)后裸鼠模型中骨肉瘤的生長情況及相關(guān)蛋白表達(dá)情況。結(jié)果表明，復(fù)方斑蝥膠囊聯(lián)合阿帕替尼可以顯著抑制骨肉瘤的生長;同時(shí)PI3K/AKT通路相關(guān)蛋白表達(dá)也隨之下降，表明該聯(lián)合用藥方案可能通過抑制PI3K/AKT通路的活性而發(fā)揮其治療作用。因此，復(fù)方斑蝥膠囊聯(lián)合阿帕替尼可能是一種可行的骨肉瘤治療方案。

關(guān)鍵詞：骨肉瘤，復(fù)方斑蝥膠囊，阿帕替尼，PI3K/AKT通路，裸鼠模型

Abstract:Thepurposeofthisstudywastoexplorethetherapeuticeffectandmechanismofcompoundhuperzinecapsulecombinedwithapatinibinthetreatmentofosteosarcoma.H&Estainingandimmunohistochemistrywereusedtoobservethegrowthofosteosarcomaandtheexpressionofrelatedproteinsinnudemousemodelsafterdrugintervention.Theresultsshowedthatcompoundhuperzinecapsulecombinedwithapatinibcouldsignificantlyinhibitthegrowthofosteosarcoma;atthesametime,theexpressionofPI3K/AKTpathwayrelatedproteinsalsodecreased,indicatingthatthiscombinationtherapymayplayatherapeuticrolebyinhibitingtheactivityofthePI3K/AKTpathway.Therefore,compoundhuperzinecapsulecombinedwithapatinibmaybeafeasibletreatmentforosteosarcoma.

Keywords:Osteosarcoma,compoundhuperzinecapsule,apatinib,PI3K/AKTpathway,nudemousemodeOsteosarcomaisararebutaggressivebonecancerthatoftenaffectschildrenandyoungadults.Despiteadvancesinchemotherapy,surgery,andradiotherapy,theprognosisofosteosarcomaremainspoor,andnewtreatmentoptionsareurgentlyneeded.

PreviousstudieshaveshownthatthePI3K/AKTpathwayplaysacriticalroleinthedevelopmentandprogressionofosteosarcoma.Activationofthispathwaycanpromotecellproliferation,migration,invasion,andtumorangiogenesis,whileinhibitionofthispathwaycaninducecellapoptosisandsensitizetumorcellstochemotherapyandradiotherapy.

HuperzineAisanaturalalkaloidextractedfromtheChineseherbHuperziaserrata.IthasbeenshowntoinhibitacetylcholinesteraseactivityandimprovecognitivefunctioninpatientswithAlzheimer'sdisease.Inaddition,huperzineAhasbeenreportedtohaveanti-cancereffectsthroughvariousmechanisms,includinginhibitingcellproliferation,inducingcellapoptosis,andsuppressingangiogenesis.

Apatinibisasmallmoleculetyrosinekinaseinhibitorthatselectivelytargetsvascularendothelialgrowthfactorreceptor-2(VEGFR-2),whichisoverexpressedinmanysolidtumors,includingosteosarcoma.Apatinibhasbeenshowntoinhibittumorangiogenesis,inducecellapoptosis,andenhancetheanti-tumoreffectsofchemotherapyandradiotherapyinpreclinicalandclinicalstudies.

Inourstudy,weinvestigatedtheanti-tumoreffectsofacombinationtherapyofcompoundhuperzinecapsuleandapatinibinnudemousemodelsofosteosarcoma.OurresultsshowedthatthiscombinationtherapysignificantlyinhibitedtumorgrowthanddecreasedtheexpressionofPI3K/AKTpathwayrelatedproteinscomparedtothecontrolandsingle-agentgroups.

ThesefindingssuggestthatcompoundhuperzinecapsulecombinedwithapatinibmaybeapromisingtherapeuticstrategyforosteosarcomabyinhibitingtheactivityofthePI3K/AKTpathwayandsuppressingtumorangiogenesis.FurtherpreclinicalandclinicalstudiesareneededtoconfirmtheseresultsandoptimizethedosingandadministrationscheduleofthiscombinationtherapyInadditiontotargetingthePI3K/AKTsignalingpathwayandsuppressingtumorangiogenesis,thecombinationtherapyofcompoundhuperzinecapsuleandapatinibmayalsohaveotherpotentialmechanismsofactionintreatingosteosarcoma.

HuperzineA,themajoractivecomponentofhuperzinecompound,hasbeenshowntohaveneuroprotectiveandanti-inflammatoryeffects.Itcaninhibittheactivityofacetylcholinesterase(AChE),anenzymethatbreaksdowntheneurotransmitteracetylcholine(ACh),andthusincreasethelevelofAChinthebrain.AChisinvolvedinvariousphysiologicalprocesses,includinglearningandmemory,andadeficiencyofAChhasbeenassociatedwithAlzheimer'sdisease.

Inadditiontoitseffectsonthenervoussystem,huperzineAhasbeenreportedtohaveanti-tumoractivityinvariouscancertypes,includingosteosarcoma.Itcaninduceapoptosis(programmedcelldeath)andinhibittheproliferationandmigrationofcancercells.Theseeffectsmaybemediatedbythesuppressionofvarioussignalingpathways,suchasthePI3K/AKT,MAPK,andNF-κBpathways.

Apatinib,ontheotherhand,isaselectiveinhibitorofvascularendothelialgrowthfactorreceptor2(VEGFR2),akeyreceptorinvolvedinangiogenesis.Angiogenesisistheprocessofcreatingnewbloodvessels,whichisnecessaryforthegrowthandmetastasisoftumors.ByinhibitingVEGFR2,apatinibcanblocktheformationofnewbloodvesselsandthusstarvethetumorofoxygenandnutrients.

Furthermore,apatinibhasbeenshowntohavedirectanti-tumoreffectsinvariouscancertypes,includingosteosarcoma.Itcaninduceapoptosisandautophagy(acellularprocessofself-degradation),inhibittheproliferationandmigrationofcancercells,andenhancethesensitivityofcancercellstochemotherapyandradiotherapy.

Takentogether,thecombinationtherapyofcompoundhuperzinecapsuleandapatinibmayhavemultiplemechanismsofactionintreatingosteosarcoma,includingtheinhibitionofthePI3K/AKTsignalingpathway,suppressionoftumorangiogenesis,inductionofapoptosisandautophagy,andenhancementofchemotherapyandradiotherapysensitivity.FurtherstudiesareneededtoelucidatetheexactmechanismsandoptimizetheclinicalapplicationofthiscombinationtherapyInadditiontothepotentialmechanismsdiscussedabove,thecombinationtherapyofcompoundhuperzinecapsuleandapatinibmayhaveothereffectsonthetumormicroenvironmentandimmunesystem.Forexample,apatinibhasbeenreportedtoinhibitthefunctionofmyeloid-derivedsuppressorcells(MDSCs)andregulatoryTcells(Tregs),whichareimmunosuppressivecellsthatplayimportantrolesinthesuppressionofanti-tumorimmunity(Zuoetal.,2019;Luetal.,2020).Ontheotherhand,huperzineAhasbeenshowntoenhancethefunctionofnaturalkiller(NK)cellsanddecreasetheexpressionofprogrammeddeath-ligand1(PD-L1)intumorcells(Lietal.,2018;Fengetal.,2020).Thesefindingssuggestthatthecombinationtherapyofcompoundhuperzinecapsuleandapatinibmayhaveimmunomodulatoryeffectsthatcontributetoitsanti-tumoractivity.

Moreover,thesafetyandtolerabilityofthecombinationtherapyshouldbeassessedinclinicaltrials.Althoughbothcompoundhuperzinecapsuleandapatinibhavebeenshowntobegenerallywell-toleratedinpreviousstudies,theircombinationmayleadtounexpectedadverseeffects.Forexample,apatinibmayincreasetheriskofbleedingandhypertension,whilehuperzineAmaycausenausea,vomiting,andothergastrointestinalsideeffects(Songetal.,2017;Huietal.,2019).Therefore,carefulmonitoringofadverseeventsiscrucialtoensurethesafetyofthepatientsreceivingthiscombinationtherapy.

Inconclusion,thecombinationtherapyofcompoundhuperzinecapsuleandapatinibholdspromiseasanoveltreatmentstrategyforosteosarcoma.Thepreclinicalstudieshaveprovidedencouragingevidenceforitsanti-tumoractivityanditspotentialsynergisticeffectswithchemotherapyandradiotherapy.However,furtherstudiesareneededtooptimizetheclinicalapplicationofthistherapyandinvestigateitssafetyandt