【doc】用蒿甲醚和吡喹酮早期治疗兔的血吸虫感染

【doc】用蒿甲醚和吡喹酮早期治疗兔的血吸虫感染 用蒿甲醚和吡喹酮早期治疗兔的血吸虫感 染 B1BL1D.ISSN0253-9756ActaPtmr~ologicaSimca中回商理1994Sepj15(5)1447--452 Earlytreatmentofschistosomalinfectionwith artemetherandpraziquantelinrabbits XIAOShu—Hua,YOUJi—Qing,MEIJin—Yan,JIAOPei—Ying (InstituteofParasiticDiseases,ChineseAcademyofPrewentiveMedicine;WHOCollabora ting CentreforMalaria,SchistosomiasisandFilariasis,Shanghai200025,China) ABSTRACTArtemether(Art,B-methyl etherofartemisinin)firstsynthesizedby ShanghaiInstituteofMateriaMedica,Chinese AcademyofSciences,iselfectiveagainstnot onlymalariabutalsoschistosomiasis.When rabbitsinfectedwithSchistosomajaponicum cercariaefor7dweretreatedigwithArt10 mg?kg-.,thetotalwormreductionrateswere 74.6—76.7.IfArt(10mg.kg)was givenonceweeklyafterthefirsttreatmentfor 3—4times,thetota1wormreductionratewas >98,andmostoftherabbitswerefree from早worms.Whenpraziquantel(Pra) wasgivenig40mg?kg一torabbitsond21af- terinfection,andrepeatedonceeveryweek for3wk,mostrabbitsshowedatotslworm reductionrate>98withtheirliversshow ingnormalormildchanges,andtheirparame— tersreIevanttoacuteschjstosomiasiswere negativeascomparedtothecontrols.Hence ArtandPraaresuggestedtobeusedinfield trialforcontrolofacuteschistosomiasisorre— ductionoftheintensityofschistosomalinfec tlon. KEYWORDSSchistosomajaponicum; artemether;praziquantel;combinationdrug therapyIdrugadministrationschedule Whenpraziquantel(Pra)wasgivento miceinfectedwithSchistosomajaponicumcer— Received199305-19Accepted1994-O6—08 TheEighthFi~-yearKeyResearchprogramofChinaN9 859170208.ProjectsupportedmpartbyJointResearch MBnrnemCommitteefromaWorldBankLoanforSchJsto- somiasis. cariaeatanearlystageafterinfection,mostor evenal1ofthe早wormsbeforesexualmaturi— tywerekilled".Whenartemether(Art, firstsynthesizedbyShanghaiInstituteofMa- teriaMedica,ChineseAcademyofSciences andfoundtobeeffectiveagainstschisto— somes'1wasgiventomiceatanearlystage afterinfection,similarresultswereob— served".Toevaluateappropriateregimens ofArtandPrausedinearlytreatmentforcon— trollingschistosomiasisandreducingthein— tensityofschistosorna1infection,thisstudy wascarriedoutinrabbits. H " oCH, Artemether MATERIALSANDMETH0DS ParasitesSchistosomajaponicumcercariae'An- huiisohte)wasreleasedfrominfectedsnailOncomvia niahupensisprovidedbytheDepartmentofVector BiologyofourInstitute. RabblteNewZealandstrainrabbits,早and, weighing2.2士0.3kg,wereprovidedbytheAnimal CareFacilitiestheInstitute.Eachrbitasin. [ectedwith198—202cercariaeviashavedabdominal skin. DrugsPrawapurchasedfromShanghai6th PharmaceuticalFactoryandArtwastheproductof KunmingPharmaceuticalFactory.PraandArtwere ?448?BIBLIDItsSN0253—9755ActaPharmacologlcaSinica中脚药理1994sep|15(5) suspended5—30mg?mlin1tragacanthandtheig volumewas2,0.kg-.. RegimensofArtandPraArtwasadmlntered igond7afterinfectionandrepeatedoncein1—2wk for3——4times.ThePratreatmentwasstartedond 21afteriraectionfollowedbyrepetitiononcein1—2 wkfor2——3times.Toevaluatetheinfluenceofad— ministrationfrequencyonefficacyofArt,rabbitsin— fectedwithcercariseweldividedinto6groups,in whichonegroupwasservedascontrolandtheother5 groupsweretreatedigwithArt10mg?kg一'ondand repeatedthedosingonceawkfor0,1,2,3and4 times. EvahmtlonofefficacyTherabbitswerekilled4 — 5wkaftertreatment.Theefficacywasevaluated bytermsofthenumberofresidualwoI;ms.Theliver wasevaluatedas一:fr?la1,freshredcolorwithout eggtubercle}土jthehverinfleshredcolotwithfew fineeggtubercles;':theliverinllghtredcolorwith someeggtubercles;++ltheliverinmarkedlyred colorwithnumeroussmalleggtubercles{+十+:the liverinmarkedgreycolorwithnumerouslargeregg tuberchs. Acuteschist~omlas'sRabbitsinfeetedwith198 — 202cercariaeweredividedintoGroupK,PandQ. TheGroupP(n=6)wastreatedigwithArt10mg ? kg一ondafterinfectionandretreatedweeklyfor4 wk.InGroupQ(n=5),Praig40mg-kgwas startedond21afterinfectinn,andrepeatedweekly for3wk.TheGroupK(=5)wasservedascon— tro1.Fourweeksaherinfection,therectaltempera— turewasmeasuredandthebloodwaswithdrawnfram theearveinforeosinophilcount.Thescbistosomal eggsinfeceswereexaminedbyhatchingandlightmi— croscope.Serumwasdetectedbycircumovalprecip— hintest(C0PT)"anddot—EL1SAusinmonoclona[ antibody(n=4ineachgroup)forschistosomalanti- bodyandcirculatingantige.,respectlvely,weekly duringthe5th一9thwkafterinfection. RESUL 1EarlytreatmentwithArtWhenArt 30mg?kg,wasgivenigtorabbitsond7af— terinfection,thetotalwormnumberwas9.0 士4.9(GroupB).whichwaslessthan127~7 ofthecontrol(GroupA).Thenumberof早 wormsinGroupBwasalsolessthanthatin thecontrol,but早wormswerestillfoundin eachrabbitofGroupB.Ingroupstreatedig withArtatthesamedoseonceawk(Group C)or2wk(GroupD)followedthefirstdos— ingfor3—4times,thetotalwormnumbersof GroupCandGroupDwerelessthanthatof GroupB.All7rabbitsinGroupCand3/7 rabbitsinGroupDwerecured(Tab1),and only1—2早wormsremainedintheother4 rabbitsofGroupD.Therabbitliversjnthe twogroupsappearedtobenorma1. WhenrabbitsweretreatedigwithArton d7afterinfectionat10orl5mg?kg,,the totalwormreductionrateswere74.6 (GroupF)and82.2(GroupG).Theliv— erchangesinGroupFandGroupGwereless apparentthanthatincontrolgroup(Tab1). Inanother3groups,therabbitsweretreated igwithArt10mg?kg(GroupH),l5rag ? kg(GroupI)or30(GroupJ)mg?kg, andrepeatedthedosingonceaweekfor4wk, thetotalwormreductionrateswere>97 (Tab1).Inthese3groups,3/8,4/8and2/ 4rabbitswerefreefrom早worm,andonlyl 一 3早wormsremainedintheotherrabbits. Meantime,noapparentchangesinthelivers weredetectedinmostoftherabbits,butin eachgroup218,2/8andl/4rabbitsshowed somefineeggtuberclesintheliverwithfresh redcolorandsoftqualityofthetissues. 2Influenceofadministrationschedule onefficacyofArtWhenArtwasgivenigto rabbitsond7(GroupL)ord7andd14 (GroupM)afterinfection.thenumbersofto— taland旱wormsofthese2groupswereless thanthoseofcontro1,but7—20早worms werefoundin4/4and2/4rabbits,respective— ly.Thetotalwormreductionratesand旦 wormreductionratesweresimilarinGroup N,OandPtreatedigwiththesamedoseof Artonceawkfor3—5times,beingmuch BIBLID.ISSN0253—9756ActsPharmacologieaSinlca中国琦理1994p{15(5,.449 Tab1.Effec忸0fartemether(Art)madpraziquant~(Pra)onrabbitsinfectedwithSchistosomajapoaicumce r 蚺r.i?.?P>O.OS,P<9.05,P<O.01vstarrespondingGroupC'J,K,P,TorY higherthanthoseo{groupLandM.Noap— parentchangesintheliverswereseeninz/4, 3/4and6/8rabbitsofthe3groups,butless andfineeggtuberclesweredetectedinthere— mainedrabbitsofthe3groups.InGroupP morecuredrabbits(5/8)wereseen(Tab1). 3EarlytreatmentwithPraRabbitsin- fectedfor21dwereinitiallytreatedigWith Pra40mg?kg-.,followedbyrepeateddosing onceaweek(GroupS)or2wk(GroupT)for 2times.InGroupT,all3rabbitswerefree {rom旱wormsandon[yafewresidual台 WOYITISwerefoundwithatotalwormreduction rateof99.5.InGroupS,thetotaland早 wormreductionrateswere87.7and 88.2,respectively,beinglowerthanthose ofGroupT(Tab1).Theliverchangesin GroupSappearedtobedarkshredcoloranda fewfineeggtubercles.Onerabbitinwhich 28residual早wormswasfoundshowedap .450-BIBLID|ISSN0253.9756}~ctaPh,trm~cblogicaSinLca中圈南理1994pI15(5) parenteggtuberclesinitsliver.InGroupT theliversof2rabbitsappevredtobenoma1. whiletheotheroneexhibitedafewoldand fineeggtubercles. WhenFrawasgiventotheinfectedrab, bitsatthesameregimenwithatota1adminis, trationtimeof3(GroupZ1)or4(GroupZ2), theefficaciesweresimilar,buttheresidual早 wornlsinGroupZ2wasIessthanthatof GroupZ1.InGroupQwiththesameregi— manofGroupZ2.thetota1and早wormre duetionrateswere98.1and99.1%,re_ spectively,and4/6rabbitswerefreefrom早 worm.Intheother2rabbits,only1or2早 wormswerefoundandnoneoronlymild changesintheliverswerenoted(Tab1). 4Pr_+Art WhenArt30mg?kg一wasgivenigtothe rabbitsond7,andPra40tug?kg一wasgiven ond21.28and35(GroupV),thenumbers oftotaland早wormswerehssthanthose treatedwithPra(GroupS)orArt(GroupU) alone(Tab1).InGroupV.3/6rabbitswere curedorfreefrom早worms,andmostlivers revealednoapparentchangeexceptfor1rab— bitwhichshowedafewfineeggtubercles. Inabovementionedcombinedtreatment thedoseofArtdecreasedto15mg?kg一 (GroupY)resultedinlessnumbersoftotal and早wormsascomplaredtoGrouxor GroupZ1.InGroupY.4/6rabbitswere cured,andintheother2rabbitsonly1resid— ua1早wormremained.Noneorverymild changesintheliverwerefound(Tab1). 5Innpence0fArtudPraontempen, ture,~eslnophilcount,andeginfeces 5.1RectaltemperatureFourwkafter infection,thetemperatureofrabbitsinCOntrol group(GroupK)was39.5?.andthengrad uallyreacheditspeakvalueof40.7?6wk 1ater.Upto9wkafterinfection.thetemper— atureofthecontrolrabbitssustainedaround 4O?-TheaveragetemperatureinGroupP andGroupQ,ie.treatedigwithArtorPra, fluctuatedbetween38.5—39.4?and38.7— 39.5?,respectively4--10wkafterinfection (FigIA). 鲻 FIg1.RtnIlteml~erature(A,a=5—6,i士)I eos|aophilCOgent(B,^=$--6,士)'dreumovs[pre- cIpJtinrate(C,_=5--6,i:)andmmtlgealevel(D, ^鲁4,士$)ininfectedlabbttstrearedI8withmrt~- meater10mll?(×)orprmdquantvl40mI? (?).Tieuntr~tcdInfectedn?1weret#mcontrol (o). 2EoslnopMIcountThanumberof eosinophilsincreasedgraduallysince5wkaf- terinfeetionandreacheditspeakvalueof (11.7土4.1)×10/L8wklater.Afterwards. thenumberofeosinophilsdecreased,butstill muchhigherthanthatbefore4wk.Noap parentalterationwasdetectedingroupstreat edwithArtorPraduring4——10wkafterin fection(Fig1B). 5.3EggsinfecesAt6—9wk,schisto— somaleggsinthefeceswerefoundincontrol group,butnotinthegroupstreatedwithArt orFra. 6I?nuencoofArt"andPrsonantibody andantigenleve1Inserllm 6.1AntibodyCOPTwasusedforde terminationofantibodyleve1.Fivewkafter infection.allrabbitsshowedpositirereaction withacircumovalprecipitinrateof33土5, andthenreachedandsustainedaround57—— 636—9wklater(Fig1C).Inallrabbits r?1J叶 O5】 :;'sli BIBLIDISSN0233—9756ActapharmacotogicaSinica中圈玮理1994SepI15(5)?451 treatedigwithArt,positivereactionrevealed in6wkafteririfectionwithacircumovalpre— cipitinrateof8,afterwards.thecircumoval precipitinratessustainedaround15—21 (Fig1C).Fiveto9wkafterinfeetion,the circumovalprecipitinratesinrabbitstreatedig withPrawere9—23(Fig1C). 6.2AntigenDot—ELISAmethodusing rnonoclonalantibodywasusedfordetermina— tionofantigen.Fiveto9wkafterinfection, theaverageserumtitreingroupstreatedwith ArtorPrawere3.4—38and2—76,respec— tively,whilethoseincontrolgroupwere45—— 1720(Fig1D). DISCUSS10N Theresultsshowedthatwhenappropriate regimenofArtorPra'wasgiventorabbits atanearlystageofinfectionwithschistosome cercariae,asignificanteffectprotectingthe hostsfromdamageinducedbyschistosome eggs,andapparentreductioninnumbersofto— taland早wormswereobserved.Itisinter- estingtonotethatthedoseofPrausedinear— lYtreatmentofrabbitswasequaltothecura— tiredoseusedinclinicaltreatment.Further studyshowedthatinexperimentallyinfected dogstheminimaleffectivedoseofPrausedat allearlystageofinfectionwasaslowas3Omg ? kg(XiaoSH,eta1.Unpublisheddata), indicatingthatthedoserecommendedforfield trialmaybebelow30mg?kg,.Sincethe doseofArtusedintheearlytreatmentofmice was300mg?kg一.higherdosesofArtwere testedinitiallyinrabbits.Unexpectedly,the effectofArt3Omg?kg一on7-day—oldschisto- somulaeinrabbitswasratherhighevenata singledose,indicatingthattheeffectofArton 7-day—oldschistosomulaeinrabbitswasmore promising.Furtherexperimentsdemonstrat— edthattheminimaleffectivedoseofArtused inearlytreatmentofrabbitswasaslowas10 mg?kg,,andtheefficacyofArtmedication commencingond7wasdependentonthefol— lowingrepeateddosingtimes. Theresultsalsodemonstratedthatwhen rabbitsweretreatedwithArtorPraatthe earlystageafterinfection,theparameterstel— evanttoacuteschistosomiasis[werenega— tire,indicatingthatmostorallofthefemale wormswerekilled.Ontheotherhand,the levelsofbothspecificantibodyandantigenin therabbitsofthetwotreatedgroupswereap— parentlylowerthanthoseofthecontrols. Therefore,theregimensofArtandPramight beusedincontrollingacuteschistosomiasis andreducingtherateandintensityofinfec— t;on. REFERENCES 1xiaoSH,YOLIJQ,MeLJY,JPY.Earlytreatmemof schistosomaILofectionwithpraziquante]Inmice. Ac姐PharmacolSin1993_14|533—8. 2LjY,YuPL.ChenYX,LiLQ,GalYZ.WangDS,et 口.S,nthesisofsomedefivative9.fartemlsinine. KexueTongbao1979'24l667—9. 3LewJ,YouJQ.YangYQ.MeLJY,GuoHF.YangHZ. ela.Studiesontheefficacyofartemetherinexperimen. taIaehhtosomjs. AetaPharmSin1982F17{187--93. 4YouJQ,MeiJY,XiaoSHEffect0artemetheragainst Schistosomajaponicum. ActaPhatraae~lS|n1992't328O一4. 8XiaoSH,Y0uJQ.MeiJY,JiaoPY.Effeet~ofearly trea廿nentofartemetheragainstschJstosomiasLscnmice. ChinJParasho[ParasiticDis1994l12|7一l2. 8D~gnosticLaboratory.fShanghaiInstituteofParasgmlc~ gy.Disnosisoschistoaomiasisjaponicabydryovapre- elptt~tlonreaction? ChLnJInternMed1979.18I197—200. 7YanzztLu2Y?WangW,WuY.XuFN.QingXY.ff at.DetectionofsehistosomeovaantigenbyDot—ELISA wjthmonoclonalantLbody. ShanghalJImmunoi1990'10|113—8. 8YueWJ.Y肌gYQ,YuSH.YuQF.Exper]meatal study0?rabbitsafterinfectionwithBchistosomhtsis. NattMedJChlr~1980l6Ot467—7O. 一 年 45Z.BIBLID=ISSN02535756ActaPlutrmacotogic~Sinlca中田药理荦报 1954Se0;15(5) 用蓠甲醴和吡哇酮早期治疗兔的血吸粤感染 肖树华,尤纪青,梅静艳,焦佩 I? 英 yS ( , 中国预防医 ~/6 再磊E而研究所,世界五生组织疟疾,血吸 虫病和丝虫病合作中心,上海200025,中国) 凸摘要兔于感染血吸虫尾蚴后d7或d21分别 开始ig1次蒿甲醚(Art)10mg?kg和吡喹酮 (Pra)40mg?kg,,然后每隔1wkig1次相同 剂量的Art或Pra连给3,4次,减旱虫率达 98以上,且部分兔无旱虫,上述兔经Art或 Pra早期治疗后,其肝脏与正常兔相仿,或仅 有轻度变化,一些与急性血吸虫病有关的指标 测定亦为阴性. 关键词日本I;莲里;堕堡联台 药物疗法}用药计划 z ………………………………………一, BIBLID:ISSN02539756ActaPharmaco[ogicaSinica中国琦理 1994sep;15(5):452--455 维拉帕米对大鼠脑突触体内ca和Ca"Mg.斗.一ATP酶的影响 —— , 哇查兰(重庆医科大学生化药理研究室,重庆630046,中国) Effectofveral~milollCaandCa Mg-ATPascactivityinratbrainsynapto- som~l DONGZhi,XUEChun—Sheng (DepartmentofBiochemicalPharmacology, ChongqingUniversityofMedicalSciences, Chongqing630046,China) ABSTRACTTo,elicitthecorrelationbetween theadrenergictransmitterreleaseandcalmod— ulin(CaM),theeffectofverapamilonthe freeCaconcentrationwasmeasuredwith fluorescenceanalysisandCaMg一 ATPaseactivityinratsynaptosomeswere studied. Whenstimulatedwithhigh—Kornore. pinephrine.theconcentrationoffreeCain rataynaptosomewasincreasedbyverapamil 10,50,and100Hmol?L_..ButthefreeCa concentrationintheresungsynaptosomewas Received1992-05-22Accepted1994-02-22 0projectSupportedhytheNatLona[NaturalScienceFouada— tionOfChina.?5880732. Greduatestudent(1591). 久j台s. reducedbyverapamil.TheactivityofCa.一 Mg"?一ATPseinsynaptosomewasremark ablyinhibitedbyverapami1inadose— dependentmanner.Theseresultssupport ourhypothesisthatCaMnotonlyactsdirectly ontheyesiclestOenhancethetransmitterre. 1ease.butalsoactsontheactivityofCa( Mg?一ATPsetOreducethefreeCainthe cytosol,andindirectlyinhibitedthetransmit— terrelease. KEYWORDScalcium:Ca…Mg- ATPase;synaptosomes}verapamil}calmod ulin;potassium;brain;norepinephrine /一摘要维拉帕米(Ver)10,50和100t-unol?L 能增加高K和去甲肾上腺素(NE)所致