化学成分半夏鱼腥草石油醚提取物的抗炎和镇痛作用精Word文件下载.docx
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化学成分半夏鱼腥草石油醚提取物的抗炎和镇痛作用精Word文件下载.docx
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FullLengthResearchPaper
Chemicalcomposition,anti-inflammatoryandanalgesicactivitiesofthepetroletherextractofPinelliacordataBaokangHuang1*,ZhiyongChu2,WeipingHe1andLupingQin1
1DepartmentofPharmacognosy,SchoolofPharmacy,SecondMilitaryMedicalUniversity,Shanghai,China.2NavalMedicalResearchInstitute,Shanghai,China.
Accepted11August,2011
Theanalgesicandanti-inflammatoryeffectofthepetroletherextractofPinelliacordata(PEPCwasinvestigatedusingthexylene-inducedearedemaandaceticacid-inducedwrithingtestinmice.PEPCwasadministeredintragastrically(i.g.at200,400and600mg/kg.Ithadsignificantanti-inflammatoryandanalgesicactivitieswithdosedependentmanner.ThechemicalcomponentsofPEPCwereanalyzedbygaschromatography-massspectrometry(GC/MS,andthemaincomponentsweremelissane(20.1%,τ-sitosterol(13.9%andlinoleicacid(10.1%.Thesedatamayaffordscientificbasisonthetraditionaluseoftheplantforpainfulandinflammatoryconditions.
Keywords:
Anti-inflammatory,analgesicactivity,Pinelliacordata,petroletherextract,gaschromatography-massspectrometry(GC/MS.
INTRODUTION
Therearemultiplereportsofnaturalproductshavingantimicrobial,anti-inflammatoryandanalgesicactivities,andtheessentialoils,sesquiterpenesandflavonoidsmightbetheiractivecomponents(Akinetal.,2010;
Lawaletal.,2010;
Luetal.,2009;
OwolabiandOmogbai,2007.InChina,manytraditionalChinesemedicineshavebeenusedtotreatpainsorinflammationcausedbyconditionssuchasarthritisandtendinitis.
Pinelliacordataisamedicinalplant(AraceaefamilynativetosoutheastChina.Theplantismarkedwitharrowheadleavesandgrowsalongforeststreamorsteeprocknearstream(Figure1.ThestemtuberofP.cordata(Chinesename“Dishuizhu”containsavarietyofconstituents,includingalkaloids,whichhaveavarietyofpharmaceuticalproperties.Thoughitisalittlepoisonous,itwastraditionallyusedasamagicalherbineastChinafortreatingallkindsofpain,stomachache,snakebitepoisoning,injury,arthritis,rheumatism,canceroustumorsandskindiseases.Capsulescontainingthiscrudedrugwereclinicallyusedasanalgesicandanti-inflammatoryagentsinZhejiangProvince(ChiangSuNewMedicinalCollege,2003;
InstituteofBotany,1979.Thepresentstudywasthereforeplannedtoinvestigatethechemical*Correspondingauthor.E-mail:
hbkcn@.Fax:
+86021-81871301.composition,analgesicandanti-inflammatoryeffectsofthepetroletherextractofP.cordata.
MATERIALSANDMETHODS
AdultICRmiceweighing18-22gwereobtainedfromAnimalCenterofSecondMilitaryMedicalUniversity.Micewerehousedincagesandmaintainedinanair-conditionedroomwithcontrolledtemperature(21±
1°
Candhumidity(55±
15%,andwerekeptona12hlight/darkcycle.Theywerefedwithstandarddietandwaterandacclimated3daysbeforetheywereused.AllproceduresinvolvinganimalswereconductedinaccordancewiththeEuropeanCommunityguidelinesfortheuseofexperimentalanimalsandapprovedbytheSecondMilitaryMedicalUniversityCommitteeonAnimalCareandUse.Theanimalsweredeprivedoffoodfor15hbeforetheexperiment,withfreeaccesstodrinkingwater.Unlessindicated,equalnumberoffemaleandmaleanimalswasusedineachexperiment.Inallexperiments,animalswererandomlydividedintogroupsoftenanimals,andeachanimalwasusedonlyonceintheexperiment.
Protocol
Eachgroupwasvehicletreated(intragastrically,i.g.,withPEPC(200,400and600mg/kg.Asareference,indomethacin(10mg/kg,i.g.(ChengduPharmaceuticalFactory,Chengdu,Chinawasusedasthestandarddrug.2%Tween20solution(TW,Merckinphysiologicalsalinewasusedasavehicle.ThedosagesofPEPCwereestimatedbasedonourpreliminaryexperimentand
11024Afr.J.Biotechnol.
recommendedoraldosageforthefolkuseinhuman.Aceticacidandxylene(SinopharmchemicalreagentCo.Ltd.,Chinawereusedastheinducerdrug.
Herbalmaterialanditsextract
P.cordatawascollectedinAugust2009fromZhejiangProvince,ChinaandwasauthenticatedbyProfessorHanchenZheng,DepartmentofPharmacognosy,SecondMilitaryMedicalUniversity.Avoucherspecimen(No.H038#wasdepositedintheherbariumoftheDepartmentofPharmacognosy,SchoolofPharmacy,SecondMilitaryMedicalUniversity.Air-driedpowderedmaterialweighing1kgwasmaceratedandsoakedinthreevolumes(w/vofpetroleumether(30to60°
Cfor24hatroomtemperature.Theprocesswasrepeatedthreetimesandfiltrateswerepooledandconcentratedat45°
Cunderreducedpressureinarotaryevaporatorandyielded22.1gofpetroleumetherextract(PEPC.PEPCwaskeptinarefrigeratorandwasfreshlydissolvedinthevehiclebeforeeachexperiment.
Gaschromatography-massspectrometry(GC/MSanalysisGC/MSanalysiswascarriedoutonaThermoFocusDSQGC–MSinstrument(Thermo,USA.Theanalyteswereseparatedona30m×
250µ
mHP-5capillarycolumncoatedwitha0.25µ
mfilmof5%phenylmethylpolysiloxane.Splitratiowas30:
1.Thecolumnoventemperaturewasprogrammedat15°
Cmin−1fromaninitialtemperatureof60to300°
C,whichwasheldfor5min.Theinjectionandionsourcetemperatureswere250°
C.Helium(99.999%wasusedascarriergasataflowrateof1ml.min−1.Theionizingelectronenergywas70eVandthemassrangescannedwas40to400amuinfull-scanacquisitionmode.CompoundswereidentifiedusingtheNISTMassSpectralSearchProgram(NationalInstituteofStandardsandTechnology,USA.
Assayofxylene-inducedinflammationinmice
Onehouraftergivingthemedicineforthelasttime,themice'
srightearwasembrocatedwithxyleneaccordingtothemethoddescribedbyliterature(Zhouetal.,2008.Theedemawasmeasured1hafterxylenetreatment,afterwhichmiceweresacrificedandaplugwasexcisedfromboththeears.Theearswellingrate(SRwasmeasuredasfollows:
SR=(WR-WL/WL,
Where,WRistheweightofright(treatedearandWListheweightoftheleft(untreatedear.Theanti-inflammatoryactivitywasexpressedasinhibitoryratio(IRorpercenttumefactionreductionintreatedmicecomparedtothecontrols.
TheIRofthetreatedgroupwascalculatedasfollows:
IR=(A-Bx100/A,
Where,AistheedemaofvehiclegroupandBistheedemaofdruggroup.Eachvaluewasthemeanofindividualdeterminationsin10mice.
Abdominalconstrictioninducedbyaceticacid
Thewrithingtestwascarriedoutfollowingthemethoddescribedbyliterature(Rabanaletal.,2005.Micewererandomlydividedintoeachgroup(n=10,andgroupsofovernightfastedmiceweretreatedwithPEPC,vehicleorindomethacin1hbeforeanintraperitonealinjectionof0.7%aceticacidat0.1ml/10gbodyweight.Thenumberofwrithingwascountedforeachanimal,starting3minafteraceticacidinjectionovertheperiodof12min.Statisticalanalysis
Resultswereexpressedasmean±
S.E.M.Statisticalanalyseswereperformedwithone-wayANOVAfollowedbyStudent'
st-test.Pvalueslessthan0.05wereconsideredtobestatisticallysignificant.RESULTSANDDISCUSSION
ThechemicalcomponentsofthepetrolextractsofP.cordatawereanalyzedbyGC/MS,and26compoundswereidentified.Themaincompoundsweremelissane(20.1%,τ-sitosterol(13.9%,linoleicacid(10.1%,nona-cosane(8.6%,hexadecanoicacid(7.7%,campesterol(6.7%andheptacosane(5.8%(Table1.Intheaceticacid-inducedwrithingtest,PEPCshowedobviousinhibitionofthewrithingresponsesinducedbyaceticacidinadose-dependentmanner,withinhibitoryratioof19.5%,42.7%and53.0%,respectively(Figure1,whiletheindomethacininduced61.6%writhinginhibition.PEPCshowedhighestinhibitoryratioatthedoseof600mg/kg.Inthewrithingresponse,aceticacidcausespainbyliberatingPGsandmanyothersthatexcitepainnerveendings(Choietal.,2006.
However,constrictioninducedbyaceticacidwasconsideredasanon-selectiveanti-nociceptivemodel,andforthechemicaldiversityofthenaturalproducts,herbextractspossessesbothperipheralandcentralanalgesicactivity(Sá
nchez-Mateoetal.,2006.ThecrudedrugofP.cordatahasbeenusedtotreatabdominalpain,waistacheandheadacheinfolktradition.InordertoclarifywhethertheactivitiesareduetotheblockadeoftheeffectorthereleaseofPGsthatexcitepainnerveendings,moreanimalmodelsincludingformalin,hotplateandtailflicktestareneededinfurtherstudy.Inthexylene-inducedearedematest,thepositivecontroldrugindomethacinat10mg/kgexhibitedsignificantinhibitoryeffect.Theinhibitionwasuptoapproximately28.7%.PEPCexertedobviousanti-inflammatoryactivityinadose-dependentmanner.Atthedoseof200,400and600mg/kg,theedemareductionswere2.6,14.2and30.8%,respectively.Atthedoseof600mg/kg,thepotencyofPEPCwascomparabletothatof10mg/kgindomethacin(Figure2.IndomethacinisaknownnonselectiveCOXinhibitor.Inxylene-inducedearedemaacuteinflammationmodel,PEPCmightactsinpartthroughtheinhibitionoftheinflammatorymediatorsoftheacutephaseofinflammationlikeindomethacin.Theresultsdemonstratetheanti-inflammatorypropertiesofPEPCandprovidedevidenceofthisherbalmedicineforthetreatmentofinflammatorydiseasesinChinese.ThePEPCisacomplexmixtureoffattyacidsandsterols,whichcomprisesagreatpartofthefraction.Itwasreportedthatstigmasterolandsitosterolcould
Huangetal.11025
Figure1.TheplantofPinelliacordataN.E.Brown.
Table
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