简介：

简介：ToinvestigatetheeffectsofoverallalkaliofatraditionalChinesemedicine“Tongbiling”(brucineandstrychninealkaloidsinmain)onthecytokinesexpressioninTh1andTh2cellsinthesynovialfluidofpatientswithrheumatismarthritisandtheirsignalpathway,themononuclearcellsinthesynovialfluid(SFMC)ofpatientswereisolatedbyFicoll-Hypaquegradientcentrifugation,andtheCD3^+CD69^+andCD3^+HLA-DRantigenwereanalyzedbyflowcytometryincomparisonwiththoseoftheperipheralblood.TherestofcellswereculturedafterresuspensionwithRPMI1640culturemedium.Phorbol12,13-dibutyrate(PDB)andionomycinwereaddedsuccessivelyintotheculturewithvariousconcentrationofoverallalkaliTongbiling(TBL).After4hofcultivation,theexpressionofIFN-γandIL-4inCD3^+cellswereanalyzedbyflowcytometry.TheinfluenceofoverallalkaliTBL(100mg/I,)ontheintracellularcalciumwasinvestigatedafterFluo-3/AMlabelingandstimulationwithPDBandionomycinat1,2,4and10min,andtheinfluenceofTBLontheexpressionofCD3^+CD69^+cellsweredeterminedwithstimulationofPDBfor24hinthewholebloodlymphocytesculture.ItwasfoundthatthepercentageofTcellsbearingCD69wassignificantlyup-regulated(77%),whilethatofTcellsbearingHLA-DRwas44%inthesynovialmononucleatedcells.AfterPDBandionomycinstimulation,theexpressionofIFN-7inCD3~cellswereup-regulated,buttherewasnochangeontheexpressionofIL-4inCD3^+cells,indicatingthatratioofTh1/Th2wassignificantlyincreasedandThcellsdifferentiatetoThlcellsinmainly.FourconcentrationsofoverallalkaloidofTBI,(200mg/L,100mg/L,50mg/L,25mg/L)coulddown-regulatedtheexpressionofIFN-γinCD3^+cellsandtheTh1/Th2ratioobviously,butalltheconcentrationsoftheoverallalkaloidshadnoeffectontheexpressionofIL-4inCD3^+cells.100mg/Lconcentrationoftheoverallalkaloiddidnotdown-regulatetheintracellularcalciumlevel.Eachconc