三七提取物对GES-1细胞及MNNG

GES-1MNNGGES-11,1,2,2,2(1.,100078;2.,100700)[]:,,():GES-1N--N--N-(N-methyl-N-nitro-N-nitroso-guanidine,MNNG)GES-1(MC);,[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide,MTT](8%4%2%)GES-1MC72h,:2h6h,8%,2h8%GES-170.8%(P0.01),2h8%MC45.3%(P0.01);6h8%GES-188.5%(P0.01),6h8%MC42.4%(P0.01):2h6hGES-1MC[];;;,[]R285.6[]A[]1672-1977(2004)06-0445-05InhibitingeffectsofPanaxnotoginsengextractsonproliferationofGES-1cellsandMNNG-transformedGES-1cellsWANGZhi-Bin1,LIJun-Xiang1,ZHULing-Qun2,NIUFu-Ling2,CUIWei2(1.DepartmentofGastroenterology,DongfangHospitalofBeijingUniversityofTraditionalChineseMedicine,Beijing100078,China;2.LaboratoryofInternalMedicine,DongzhimenHospital,BeijingUniversityofTraditionalChineseMedicine,Beijing100700,China)ABSTRACTObjective:Throughcellcultivation,westudiedtheinhibitingeffectsoftheserumofthedogfedwithPanaxnotoginsengextractsonprecancerousgastriccells,tryingtofindthebesttimepointsorperiodswhentheextractsfunctionwasthestrongestafteradministrationoftheextractstothedog.Methods:TheexperimentsadoptedeternalizedhumangastricmucosaepitheliumGES-1cellsandMCcellsgainedfromGES-1cellstransformedbyN-methyl-N-nitro-N-nitroso-guanidine(MNNG)asthemodelofprecancerouslesionsforstudyinvitro.WetooktheserumofadogbeforeandatdifferentpointsoftimeafterfeedingthedogwithPanaxnotoginsengextractsforexperiment.BymeansofMTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide)assay,weexaminedtheinhibitingeffectsoftheserumafterculturingtheGES-1andMCcellsfor72hourswithdifferentconcentration(8%,4%,2%)ofmedicatedserumobtainedfromthedogatdifferentpointsoftime,soastofindthat,atwhichpointsoftimethemedicatedserumobtained,itwouldbethemosteffective.Results:TheresultsshowedthattheGES-1andMCcellsinhibitionratesofmedicatedserumfromthepointsof2-hourand6-hourwerethehighest,andtheculturemediumcontaining8%ofmedicatedserumfromthesetwopointshadprominentinhibitingeffectsonbothkindsofcells.TheGES-1cellsinhibitionrateinculturemediumcontaining8%ofmedicatedserumfromthepointof2-hourwas70.8%(P0.01)andthatoftheMCcellswas45.3%(P0.01).TheGES-1cellsinhibitionrateinculturemediumcontaining8%ofmedicatedserumfromthepointof6-hourwas88.5%(P0.01)andthatoftheMCcellswas42.4%(P0.01).Conclusion:Thepointsoftimewiththestrongestinhibitingeffectsare2hoursand6hoursafterbeingfedwithPanaxnotoginsengextracts.Atthesetwopoints,theserumismosteffectiveininhibitingtheproliferationofGES-1andMCcells.KEYWORDSPanaxnotoginseng;gastriccancer;cellculture;animals,laboratoryJChinIntegrMed,2004,2(6):445-449,54420041126JChinIntegrMed,Nov2004,Vol.2,No.6[](No.2024)[](1978-),,,.E-mail:[e-mail]:Prof.LIJun-Xiang.E-mail:[e-mail][1],,,[2,3],GES-1N--N--N-(N-methyl-N-nitro-N-nitroso-guanidine,MNNG)GES-1[4](MC),,,()11.11.1.1GES-1MC1.1.2(10kg,100L,2h,80L,1.5h,2,50L,95%50L,24h,,,70,)1.1.3DMEM(DulbeccosmodifiedEaglesmedium)/,Gibco[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide,MTT],Sigma;(),;2496,Nunc1.1.4CO2,NAPCO5410;,ThermoLabsystemDSXSystem;,OlympusIM21.21.2.11,,0.16g/kg,,5103060min234568101214162024h,-201.2.2:DMEM,3.6g/LNaHCO3,0.22m,4:90%10%,10U/L1.2.3GES-1MC375%CO2(80%90%),131.2.4GES-1MC,8%4%2%GES-1MC,0.1%0.1%(ethylenediaminetetraaceticacid,EDTA),4104/ml,96,0.1ml24h,,72h,1.31.3.1GES-1MC1.3.2GES-1MC,,MTT(5mg/ml)20l,0.1ml,4h,(dime-thysufoxide,DMSO)100l,10min540nmOD:=(OD-OD)/OD100%1.4SPSS10.0,studentst22.1GES-1,GES-1,,,,(1),2h4%,GES-1,,,,(2)GES-1,5min4%5min2%10min2%30min4%30min2%8h8%16h2%,(P0.05P0.01)64420041126JChinIntegrMed,Nov2004,Vol.2,No.62h6hGES-1,2h8%70.8%(P0.01),2h4%47.4%(P0.01);6h8%88.5%(P0.01),6h4%63.5%(P0.01)12.2MC,MC,,MC,,,;,,,,(3),2h4%,MC,,,,(4)MC,5min8%5min4%5min2%10min2%30min8%30min2%1h4%2h2%3h4%5h4%6h4%10h4%12h2%14h2%16h2%,(P0.05P0.01)2h6hMC,2h8%45.3%,2h4%8.6%(P0.05);6h8%42.4%(P0.01),6h4%4.6%(P0.01)2172hGES-1(200)Fig1GES-1cellscultivatedfor72hoursinculturemediumcontainingnormalserum(200)24%(2h)72hGES-1(100)Fig2GES-1cellscultivatedfor72hoursinculturemediumcontaining4%ofmedicatedserumfromthepointof2-hour(100)1GES-1Tab1InhibitingeffectsofmedicatedserumonGES-1cells(xs,%)TimepointserumobtainedfromthedognInhibitionrateinculturemediumcontainingdifferentconcentrationofserum8%ofserum4%ofserum2%ofserumBeforeadministrationofextractstothedog721.37.0-4].63.5-26¥.14.25minafteradministration728.63.6*-6].65.9-28¥.27.310minafteradministration739.31.7**-18].73.8**-26¥.13.430minafteradministration733.410.2*-6].35.4-23¥.54.81hafteradministration754.53.0**34].39.2**1¥.44.2**2hafteradministration770.83.0**47].43.4**-0¥.84.8**3hafteradministration755.13.7**47].46.9**19¥.35.6**4hafteradministration752.32.9**20].56.2**16¥.32.8**5hafteradministration779.43.2**63].75.2**36¥.33.3**6hafteradministration788.51.5**63].53.8**41¥.23.2**8hafteradministration743.66.99].54.7**18¥.17.1**10hafteradministration7-16.17.3**-52].510.5**10¥.05.7**12hafteradministration7-13.34.8**-35].74.5**3¥.47.7**14hafteradministration7-36.512.1**-58].012.2**10¥.15.7**16hafteradministration72.74.8**-20].95.1**24¥.64.320hafteradministration714.