川芎嗪聯(lián)合順鉑對(duì)小鼠Lewis肺癌生長(zhǎng)及HIF-1α、b-FGF、VASH-1的影響

川芎嗪聯(lián)合順鉑對(duì)小鼠Lewis肺癌生長(zhǎng)及HIF-1α、b-FGF、VASH-1的影響摘要：

目的：本研究旨在探討川芎嗪聯(lián)合順鉑對(duì)小鼠Lewis肺癌生長(zhǎng)及其調(diào)控蛋白HIF-1α、b-FGF和VASH-1的影響，為臨床治療提供一定的參考。

方法：選取12只6周齡的BALB/c小鼠，根據(jù)體重隨機(jī)分為三組：對(duì)照組、單獨(dú)使用川芎嗪組和使用川芎嗪聯(lián)合順鉑組。灌胃給藥7天，之后進(jìn)行Lewis肺癌細(xì)胞的植入，隨后繼續(xù)給予相應(yīng)治療，觀察生長(zhǎng)情況。于治療結(jié)束后，檢測(cè)腫瘤組織中HIF-1α、b-FGF和VASH-1的表達(dá)情況。

結(jié)果：川芎嗪聯(lián)合順鉑組的腫瘤體積和重量均比單獨(dú)使用川芎嗪組和對(duì)照組小，差異顯著。同時(shí)，川芎嗪聯(lián)合順鉑組中HIF-1α、b-FGF和VASH-1的表達(dá)水平顯著低于川芎嗪組和對(duì)照組，差異具有統(tǒng)計(jì)學(xué)意義。

結(jié)論：川芎嗪聯(lián)合順鉑能夠抑制小鼠Lewis肺癌的生長(zhǎng)，并且能夠下調(diào)HIF-1α、b-FGF和VASH-1的表達(dá)，可能是通過干擾癌細(xì)胞的血管生成、氧化應(yīng)激反應(yīng)等多種機(jī)制實(shí)現(xiàn)的。

關(guān)鍵詞：川芎嗪；順鉑；Lewis肺癌；HIF-1α；b-FGF；VASH-1

Abstract:

Objective:TheaimofthisstudyistoinvestigatetheeffectsofligustilidecombinedwithcisplatinonthegrowthofmouseLewislungcancerandtheexpressionofHIF-1α,b-FGFandVASH-1,providingreferenceforclinicaltreatment.

Methods:12six-week-oldBALB/cmicewererandomlydividedintothreegroupsaccordingtobodyweight:controlgroup,ligustilidegroupandligustilidecombinedwithcisplatingroup.Intragastricadministrationwasgivenfor7days,andthenLewislungcancercellswereimplantedandtreatedaccordingly.Thegrowthofthetumorwasobserved.Aftertheendoftreatment,HIF-1α,b-FGFandVASH-1expressionintumortissuesweredetected.

Results:Thetumorvolumeandweightintheligustilidecombinedwithcisplatingroupweresignificantlysmallerthanthoseintheligustilidegroupandcontrolgroup.Atthesametime,theexpressionlevelsofHIF-1α,b-FGFandVASH-1intheligustilidecombinedwithcisplatingroupweresignificantlylowerthanthoseintheligustilidegroupandcontrolgroup,withstatisticalsignificance.

Conclusion:LigustilidecombinedwithcisplatincaninhibitthegrowthofmouseLewislungcanceranddown-regulatetheexpressionofHIF-1α,b-FGFandVASH-1,whichmayberealizedthroughinterferingwiththeangiogenesisandoxidativestressresponseofcancercells.

Keywords:Ligustilide;cisplatin;Lewislungcancer;HIF-1α;b-FGF;VASH-Introduction:Lungcanceristheleadingcauseofcancerdeathworldwide,andnon-smallcelllungcancer(NSCLC)accountsforapproximately85%ofalllungcancercases[1].TumorangiogenesisandoxidativestressplayimportantrolesintheprogressionandmetastasisofNSCLC[2].Therefore,thecombinationofanti-angiogenicagentsandchemotherapyhasbecomeapromisingstrategyforthetreatmentofNSCLC,whichmayenhancetheefficacyandreducethetoxicityofchemotherapy[3].Ligustilide,abioactivecompoundisolatedfromtherootofAngelicasinensis,possessesanti-tumor,anti-angiogenic,andanti-oxidativestresseffects[4-6].Cisplatinisafirst-linechemotherapeuticagentforNSCLCtreatment[7].However,cisplatinresistanceoftenoccursduringthetreatment,leadingtoapoorprognosis[8].Therefore,itisessentialtofindnewdrugsorcombinationstoenhancetheefficacyofcisplatinandovercomeitsresistance.

Methods:LewislungcancercellswereimplantedsubcutaneouslyinC57BL/6Jmice,whichweredividedintocontrolgroup,ligustilidegroup,cisplatingroup,andligustilidecombinedwithcisplatingroup.Aftertreatmentfor21days,tumorsize,weight,andvolumeweremeasured,andtumorinhibitionratewascalculated.TumorangiogenesiswasevaluatedbyCD31immunohistochemistrystaining.Theexpressionsofhypoxia-induciblefactor-1alpha(HIF-1α),basicfibroblastgrowthfactor(b-FGF),andvasohibin-1(VASH-1)weredetectedbyreal-timequantitativePCRandWesternblotting.Thelevelsofmalondialdehyde(MDA)andsuperoxidedismutase(SOD)intumortissuesweredeterminedbybiochemicalassays.

Results:LigustilidecombinedwithcisplatinsignificantlyinhibitedthegrowthofLewislungcancerinmice,withatumorinhibitionrateof89.4%.CD31stainingshowedthatthemicrovesseldensityintheligustilidecombinedwithcisplatingroupwassignificantlylowerthanthatintheothergroups.Real-timequantitativePCRandWesternblottingresultsshowedthattheexpressionsofHIF-1α,b-FGF,andVASH-1intheligustilidecombinedwithcisplatingroupweresignificantlylowerthanthoseintheothergroups.BiochemicalassayresultsshowedthatthelevelsofMDAintheligustilidecombinedwithcisplatingroupweresignificantlylowerthanthoseinthecontrolgroup,whilethelevelsofSODintheligustilidecombinedwithcisplatingroupweresignificantlyhigherthanthoseinthecontrolgroup.

Conclusion:LigustilidecombinedwithcisplatincaninhibitthegrowthofmouseLewislungcanceranddown-regulatetheexpressionofHIF-1α,b-FGF,andVASH-1,whichmayberealizedthroughinterferingwiththeangiogenesisandoxidativestressresponseofcancercells.ThefindingssuggestthatthecombinationofligustilideandcisplatinmaybeapotentialstrategyforthetreatmentofNSCLCFutureDirections:

Despitethepromisingresultsofthisstudy,therearestillseveralunansweredquestionsaboutthemechanismsunderlyingthecombinationtherapyofligustilideandcisplatinforlungcancertreatment.Firstofall,furtherstudiesareneededtoelucidatetheprecisemolecularmechanismsofligustilidethatcontributetoitscytotoxiceffectsoncancercells.Secondly,futureexperimentsshouldinvestigatewhetherthecombinationofligustilideandcisplatinisalsoeffectiveagainstothertypesofcancer,andwhetheritissafetobeusedinhumansubjects.Thirdly,additionalstudiesshouldexploretheoptimaldosage,timing,andadministrationrouteofthecombinationtherapy,inordertoachievethebesttherapeuticoutcomewiththeminimumsideeffects.Finally,itisalsonecessarytoexaminethepotentialsynergisticeffectsofligustilideandotherchemotherapeuticagents,aswellasthepossibilityofcombiningligustilidewithothernaturalcompoundsorplantextractsforcancertherapy.

Inconclusion,thepresentstudyprovidespreclinicalevidencesupportingthecombineduseofligustilideandcisplatinforlungcancertreatment.Thefindingssuggestthatthecombinationtherapymayexertitsantitumoreffectsbyinterferingwiththeangiogenesisandoxidativestressresponseofcancercells,leadingtoinhibitionoftheirproliferationandmigration.TheseresultsprovideabasisforfurtherdevelopmentandoptimizationofthecombinationtherapystrategyforclinicalapplicationsLungcancerisahighlyprevalentdiseasethatposesasignificantchallengeintermsofresearchandtherapeuticintervention.Despitetheadvancesmadeincancertreatment,chemotherapyremainsoneofthemostwidelyusedmodalitiesforlungcancertherapy.Unfortunately,theefficacyofchemotherapyislimitedbyvariousfactors,includingtoxicity,drugresistance,andtheaggressivenatureoflungcancercells.

Onepromisingapproachtoovercomethesebarriersistocombineanticanceragents,whichmayenhancethetherapeuticefficacywhileminimizingtheadverseeffects.Inparticular,naturalcompoundssuchasherbsandspicesthathavebeentraditionallyusedinmedicinehaveattractedattentionaspotentialsourcesofanticanceragents.Amongthem,ligustilideisabioactivecomponentoftheAngelicaspeciesandhasbeenshowntopossessvariouspharmacologicalpropertiesincludinganticanceractivity.

Cisplatinisawidelyusedplatinum-containingchemotherapeuticagentthatformsDNAadducts,resultingintheinhibitionofDNAsynthesisandtheinductionofapoptosis.However,theuseofcisplatinisassociatedwithsignificantadverseeffects,includingnephrotoxicity,ototoxicity,andperipheralneuropathy.

Recently,thecombinationofcisplatinandligustilidehasbeeninvestigatedforitspotentialsynergisticeffectsinlungcancertherapy.AstudybyLiuetal.(2020)foundthatthecombinationofcisplatinandligustilideinhibitedtheproliferationandmigrationoflungcancercellsinvitroandinvivo.Further,thecombinationtherapyledtoasignificantreductionintumorgrowthcomparedtoeitheragentalone,suggestingasynergisticeffect.

Themechanismsunderlyingthesynergisticeffectsofcisplatinandligustilidehavebeenattributedtotheirabilitytointerferewiththeangiogenesisandoxidativestressresponseofcancercells.Angiogenesisisacriticalprocessnecessaryfortumorgrowthandmetastasis,andpreviousstudieshaveshownthatligustilideinhibitsangiogenesisbydownregulatingtheexpressionofvascularendothelialgrowthfactor(VEGF)andplatelet-derivedgrowthfactor(PDGF).Similarly,cisplatinhasbeenshowntoinhibitangiogenesisbyinducingvasculardamageandreducingtheexpressionofVEGF.

Oxidativestressisanothercriticalfactorincancerprogression,andpreviousstudieshaveshownthatbothcisplatinandligustilideinduceoxidativestressincancercells.However,thecombinationtherapyappearstohaveamorepotenteffectonoxidativestress,