中药药理与临床

3,33,,,,(,221004):(GBE):MTT;HPLC;T2SODMDA:5,4GBE,2hGBE;20%(MC);GBE:GBE,GBE;;;HPLC[13],,,,(Ginkgobilobaextract,GBE)(DiabeticNephropathy,DN)[4],,(Mesangialcells,MC),HPLCMTT,,GBE,GBE,T2SODMDA,11.1GBE(No.040029),),GBE,0.5%CMC2Na1.2SD,SPF,210230g,,1.3DMEM:Gibco(withD2glucoseat25mmol/L);(MTT):Sigma(No.M2128);:(No.061221);:Gibco(No.27250018);SODMDA:(No.20080628)1.4class100CO2:Sheldon;270:Form;CK4:O2LYMPUS;128ce:Clinibio;Shimad2zuLC210A:(SCL210Avp,SPD2M20A,DGU220A,LC210ADvp,7725i)1.51.5.1GBE324,GBE(100200400mg/kg),CMC2Na,,4,,5630min,0.22m,2701.5.2MCMC10%DMEM(100U/ml),375%CO21.5.3,MC11069624h,041020304060100160200l,10%DMEM,200l,025101520305080100%,648h,,MTT(5mg/L)20l,4h,150l,,15min,A4921.5.4GBE54,9,6,400mg/kg,2(),4,15306090120150180210240min,3000rpm10min,,220,5630min,0.22m,2700.5ml,2(114)3ml,2min,4000rpm10min,,1.5ml,2min,4000rpm10min,,,35N2200l20.1%(80:20),14000rpm15min,,kromasilC18(,250mm4.6mm,5m);A,0.1%B,1;:1.0ml/min;:350nm;:20l472009;25(3)3,:07KJD360225331(min)A(%)B(%)053035706554035406560405040456055507045505550HPLC,,1.5.5GBET2SODMDAMC24h,40l,10%DMEM,200l,648h,,1.0106/ml,1000rpm3min,PBS,4400lPBS,,T2SODMDA1.5.61.5.1,MC1.5.224h,40l,10%DMEM,200l,200l10%DMEM,648h,1.5.5T2SODMDA22.12.1.12,10%,2%5%10%15%20%;30%50%;80%100%,,20%2(xs,n=6)(%)MTT(OD492nm)100.750.06220.750.13350.780.104100.820.155150.810.086200.800.087300.870.1738500.920.12339800.400.0833101000.340.06330%3P0.05,33P0.012.1.2GBE3,GBE120minGBE,GBE3GBE(xs,n=6)(min)151578721473026946211260342796146903570132241206152811563315035800322518022574518821019299917324012391916233P0.012.1.3GBET2SODMDA4,GBEMCT2SODMDA,120minT2SODMDA,GBE4GBET2SODMDA(xs,n=6)(min)SOD(U/mgprot)MDA(nmol/mgprot)1158.8150.136.540.242309.0740.156.540.203609.2960.106.150.2549011.3330.175.380.21512016.8890.28333.850.2333615012.5930.195.150.12718011.6520.185.960.18821010.8380.166.080.1992409.6670.176.130.2133P0.012.2GBET2SODMDA5,T2SODMDA,GBE,GBE5GBET2SODMDA(xs,n=6)(mg/kg)SOD(U/mgprot)MDA(nmol/mgprot)09.070.156.540.20GBE1009.2960.106.150.25GBE20011.330.1735.380.213GBE40016.890.28333.850.23333P0.05,33P0.0133.1,,572009;25(3)[5],2,3d,1h[6][7],,,,HPLCGBE,,,3.2[8],,[9],,20%MTTMC,,10%,30%50%MC,80%100%MC,20%MC,,20%,GBE3.3,,[10][11]:=,Lu[12]GBE,80mg/kg,,20%,400mg/kg3.4GBEGBEMC,MCT2SODMDA,GBE200mg/kg400mg/kgMCT2SODMDA,GBE,Yang[13],GBEGBEHPLCMTTGBE,,GBEGBE,1IwamaH,AmagayaS,OgiharaY.Effectofshosaikoto,aJapanaeseandChineseherbalmedicinalmixture,onthemitogenticactivityofLipopolysac2charideAnewpharmacologicaltestingmethod.Ethmopharmacol,1987;21(1)45532.22.TDM,1988;(5)543,,,..,2004;24(8)7414HainesDD,BakL,FerdinandyP,etal.CardioprotectiveeffectsofthecalcineurininhibitorFK506andthePAFreceptorantagonistandfreeradicalscavenger,EGB761,inisolatedischemic/reperfusedrathears.JCardioyascPharmacol,2000;3537445,,,..,1996;3(3)126,..,1999;15(6)5695707,..,2000;24(2)89928,,,..,2000;16(1)1181199,,,..2,2002;4(3)515510,..,2007;27(3)788011,,,..,1997;13(3)293112LuQ,YinXX,WangJY,etal.EffectsofGinkgobilobaonpreventionofdevelopmentofexperimentaldiabeticnephropathyinrats.ActaPharmacolSin,2007;28(6)81882813YangXF,WangNP,LuWH,etal.EffectsofGinkgobilobaextractandtanshinoneoncytochromeP2450isozymesandglutathionetransferaseinrats.ActaPharmacolSin,2003;24(10)10331038EstablishmentandvalidationofserumpharmacologymethodforGinkgobilobaextractWeiYaqin,TangDaoquan,GaoYuanyuan,YinXiaoxing,ChenYonggang,HanLin(XuzhouMedicalCollege,Xuzhou221004)Objective:ToestablishandvalidatedaserumpharmacologymethodforGinkgobilobaextract(GBE).Methods:MTTassaywasusedtodeterminethesuitableconcentrationofblankseruminculturesystem.HPLCmethodwasusetodeterminetheoptimumdosageofadminis2trationandtimeofhemospasia.T2SODandMDAwasselectedtovalidatetheantioxidanteffectofserumcontainingGBE.Results:RatswereoraladministrationofGBEby5timesdoseofadultstwiceaday,keepingon4days.Hemospasiaafter2hoursofthelastadministrationwasthebestpreparativemethodoftheserumcontainingdrug.Adding20%ofthisserumtoculturesystemhasnoinfluenceonmultiplicationof672009;25(3)mesangialcells(MC).ThisserumcontainingGBEhasobviousantioxidanteffectonMC.Conclusion:SerumcontainingGBEpreparedbysuitablemethodpossessesobviousantioxidanteffectandthisserumpharmacologymethodcanbeappliedtoevaluatethepharmacologicaleffectandmechanismofGBE.KeywordsGinkgobilobaextract;antioxidant;HPLC;serumpharmacology31,133,2(1,610041;2,625000),,,,,;;,,,,,,,,,,,,,200,,1,,,;,,,,20,[1,2],[3],,[4],:,:,:,::,,PolygonummultiflorumThunb.(Sect.Tiniaria),1998(FallopiaAda2ns.),20,7,Fallopiamultiflora(Thunb.)Harald.,[5,6],[7],CynanchumbungeiDecne.,,2[7,811],2.12,3,5,4’2222O22D()3,5,4’2()3,5,4’2232O2D()()2005,1.0%,0.70%2.20.3%0.8%,282O22D[12],,2.3,0.15%0.30%,,,40%55%2.4772009;25(3)3[](973)(2009CB522801)33