47-种中药对酪氨酸酶活性的影响及酶动力学的研究

:(972290):1,,1160272,,1160213,,1160274,,1160274712334[]:47::19,();:,[];;;Theeffectsof47traditionalChineseherbsontheactivityoftyrosinaseandenzymekineticsTuCaixia,LiuZhili,RenFeng,etal.DepartmentofDermatology,the2ndAffiliatedHospitalofDalianMedicalUniversity,Dalian,116027[Abstract]Objective:Toevaluatetheeffectsof47traditionalChineseherbs,frequentlyusedtotreatvitiligo,ontheactivityoftyrosinase.Methods:Experimentsinvitrowereperformedtoevaluatetheeffectsoftheethanolex2tractsfromthetraditionalChineseherbsontheactivityoftyrosinaseandenzymekinetics.Results:Theresultsshowedthat19extractscouldactivatetyrosinase,amongthemtheactivatingeffectsoftheextractsfromCaulisSpatholobussuberectus(Jixueteng),Prunellavulgaris(Xiakucao),FructusLigustrilucidi(Nvzhenzi),HerbaMen2thae(Bohe),SemenAstragalicomplanati(Tongjili),RhizomaDrynariae(Shenjiang),HerbaEcliptae(Hanlian2cao),RadixScutellariae(Huangqin),HerbaLycopi(Zelan),RadixGlycyrrhizae(Gancao)andPseudobulbuscre2mastraeappendiculataediculatae(Maocigu)weresignificantlymorestrongthanthatofpsoralen.EnzymekineticanalysisindicatedthatethanolextractsfromthetraditionalChineseherbscouldinduceacompetitive,noncompetitiveormixedactivationontyrosinase.Conclusion:TheeffectivenessoftraditionalChinesemedicineforvitiligomayberelatedtotheiractivationontyrosinase.Theherbsinducingmixedactivationappearmorepotentialinactivatingty2rosinase.[Keywords]traditionalChinesemedicine;tyrosinase;activation;kinetics,,,,,,1,23,4711.11.1.1L-(Sigma);(Sigma);();1.1.2UV754()1.1.3,,,,,,,,,,,,,,,,,,,,,,,,,,,654ChinaJLeprSkinDis.Jun2006,Vol.22,No.6,,,,,,,,,,,,,,,,,,,,1.21.2.180,(20)5g,90%50100mL,,1,90%1,23000rPmin,10min,,,,1mL41.2.2[4],1P15molPL,pH6.80.5mL100L(60u),2510min,3molPLL-2mL,2min,475nmA475,100mgPmL(mL100mg):(%)=[(C-D)-(A-B)]P(A-B)100,A=A475-+;B=A475--;C=A475++;D=A475+-,5mgPmL1.2.3[5],0.5mL(1P15molPL,pH6.8),100L(60u),2510min,,2min,475nmA475,475nm3.7103,,Lineweaver-Burk,Km1.3t22.147,19,12.2:,,,,Km,,Km,,,,,Km,192119(€xs)%286.115.56386.506.373243.1618.35366.6720.44150.006.52364.3512.05149.2424.43343.727.88142.0314.49342.6217.23135.766.70340.246.45117.585.50326.335.18113.7318.87321.9412.94101.4223.61321.182.89100.0011.463(500gPmL)47.087.7833(P0.05)2(€xs)(%)117.6101.4100.026.3321.921.263.2243.2149.2113.786.566.740.299.8286.1150.0142.0135.864.443.742.6135.83,6,7,891047,19,,11,,19,11(P0.05),7542006622612,5--8-12,,,,13,14,:,,(2),1CarlieG,NtusiNB,HulleyPA,etal.KUVA(khellinplusultravi2oletA)stimulatesproliferationandmelanogenesisinnormalhumanmelanocytesandmelanomacellsinvitro.BrJDermatol2003;149:707-717.2HearingVJ,JimenezM.Mammaliantyrosinasethecriticalregulato2rycontrolpointinmelanocytepigmentation.IntJBiochem1987;19:1141-1147.3..:,1990.525-538.4MatsudaH,HigashinoM,NakaiY,etal.Studiesofcuticledrugsfromnaturalsources.IV.InhibitoryeffectsofsomeArctostaphylosplantsonmelaninbiosynthesis.BiolPharmBull1996;19:153-156.5MaedaK,FukudaM.Arbutin:mechanismofitsdepigmentingac2tioninhumanmelanocyteculture.JPharmacolExpTher1996;276:765-769.6MarwanMM,JiangJW,deLauroCastrucciAM,etal.Psoralensstimulatemousemelanocyteandmelanomatyrosinaseactivityintheabsenceofultravioletlight.PigmentCellRes1990;3:214-221.7ChakrabortyDP,RoyS,ChakrabortyAK.Vitiligo,psoralen,andmelanogenesis:someobservationsandunderstanding.PigmentCellRes1996;9:107-116.8EspinJC,WichersHJ.Activationofalatentmushroom(Agaricusbisporus)tyrosinaseisoformbysodiumdodecylsulfate(SDS).Ki2neticpropertiesoftheSDS-activatedisoform.JAgricFoodChem1999;47:3518-3525.9EspinJC,WichersHJ.Kineticsofactivationoflatentmushroom(Agaricusbisporus)tyrosinasebybenzylalcohol.JAgricFoodChem1999;47:3503-3508.10EspinJC,vanLeeuwenJ,WichersHJ.Kineticstudyoftheacti2vationprocessofalatentmushroom(Agaricusbisporus)tyrosinasebyserineproteases.JAgricFoodChem1999;47:3509-3517.11SehgalVN.Acomparativeclinicalevaluationoftrimethylpsoralen,psoralenand8-methoxypsoralenintreatingvitiligo.IntJDerma2tol1975;14:205-208.12..:,1983.547,795.13NomenclatureCommitteeoftheInternationalUnionofBiochemistry(NC-IUB).Symbolismandterminologyinenzymekinetics.Rec2ommendations1981.EurJBiochem1982;128:281-291.14DixonM,WebbEC.Enzymes.3rdedition.NewYork:AcademicPressInc1979.381-395.(:2005-10-24:2006-01-04)1,73,5,,4,,1,3,,::,112500:2cm1.5cm,,,,,4,,:,0.9%250mL150mgC5.00.9%500mL0.5,1,500mL24U,2,1h,10mgqd,3,10%10mL,bid,,6,5,,,,,,10%(:2006-02-23)854ChinaJLeprSkinDis.Jun2006,Vol.22,No.6