肺積寧方誘導人肺腺癌A549細胞凋亡及下調VEGF表達的實驗研究.pdf

1、遼寧中醫(yī)藥大學碩士學位論文肺積寧方誘導人肺腺癌A549細胞凋亡及下調VEGF表達的實驗研究姓名：張巍申請學位級別：碩士專業(yè)：中西醫(yī)結合臨床指導教師：殷東風20080530FeiJiNingDecotioninducedapoptosiS’anddownregulation●r、，、n’1o111’eXDreSSlOnofV匕bp1nhUmanlungaOenocarclnomace上上上1neA549：anexperimentalstu

2、dyZhangWeiYinDong—fengDepartmentofInternalOncology，Affi1iatedHospitalofLiaoningUniversityoftraditionalChinesemedicineAbstractObject：ToexploretheeffectofFeiJiNingDecotion(FJN)extractSonproliferation、apoptosiSandexpression

3、ofVascularendothelialgrowthfactor(VEGF)inhumanlungadenocarcinomacell1ineA549Method：CultureA549cellinvitro，theproliferateinhibitratewasevaluatedbyMTTassay，ApoptotiCratewasdeterminedPI／AnnexinV—FITCdoublestaining，themorpho

4、logicalchangeofapoptoticcellwasobservedbyfluorescencemicroscopeandthegrowthstateofcellwasobservedbyinvertedmicroscopetheexpressionofVEGFwasdetectedbyELISAResults：MTTassayshowedthatvariousdoseteamscaninhibithumanlungadeno

5、carcinomaA549proliferationinadosedependentmannerTheIC50iS200mg／ml，weselectedfourdifferentConcentra—tion(30mg／ml，l00mg／ml，200mg／ml，1lOOmg／m1)aSexperimentalconcentration，andexperimentaltimeiS72hMorphologicalstudyshowedthat

6、，withthedoseofincreasing，thecelladhesionabilitydecreased，manycellsshedfromtheculturebottlewallandfloatinginculturemediumTheconfigurationchangestheCircle，thevolumereduces，thecel1spacingincreases，andlightrefr鹋tionisstronge

7、rAnnexinV／PIdoublestaining，ThecellapoptosiSwasevaluatedbyflowcytometryitshowsthatFJNdecotioncandeducecellapoptosiS，whichincreasewithraisedconcentrationAfterthechisquaretest，thecellapoptosiSoffourdoseteamiShigherthancontr

8、olgroup，thedifferenceiSremarkable(PO05)：thehighdoseteam(1100mg／m1)iShigherthancontrolgroup(43％)obviously，pO01AnnexinV／PIdoublestaining，atypicalsetofchangessuchascellshrinkage，cleavageofnucIear，chromatincondensation，membr

9、aneblendingandapoptoticbodyformationwillbeobservedinfluorescencemicrosxopeA11doseteamcandownregulationVEGFexpression，andthedegreesoftheirdeclineswereinadose—dependentfashionwhentheConcentrationiS200mg／mlandllOOmg／ml，thdi