凝血酶對(duì)缺氧-復(fù)氧As的影響及其與iNOS、NO的關(guān)系.pdf

1、中南大學(xué)碩士學(xué)位論文凝血酶對(duì)缺氧復(fù)氧As的影響及其與iNOS、NO的關(guān)系姓名：楊麗娟申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：生理學(xué)指導(dǎo)教師：鄔力祥20060501ABSTRACTobjectiveObservetheeffectsofthrombinonculturedastrocyteandexploreitsrelationshipwithinduciblenitricoxidesynthase(iNos)andnitricoxide(NO)Met

2、hodsPrimaryastrocytewasculturedinDN匝Mwith10％15％calfserumThecelldamageandviabilityweredetectedbythenitriteassayandthe3(4，5dimethylthazol2y1)2，5diphenyltetrazoliumbromide(MTT)conversionmethodTheflowcytometryWasperformedtoe

3、valuatetheapoptosisrateofastrocyteiNOSmRNAWasexaminedbysemi—quantitativereversetranscriptionpolymerasechainreaction(RTPCR)ImmunocytochemistryWasusedtoobservetheexpressionofiNOSproteinResults①Effectofthrombin(25，5，10，20，4

4、0U／，m1)onastrocytewithouthypoxia／reoxygenationffI／R)WasevaluatedWhencomparingopticaldensity(oD)valuesandNOproductionofdrugtreatedandcontrolcells，theeffectWassignificantonlyatthehigherconcentrationofTB(2040U／m1)used(P005)

5、②TheproductionofNOWasincreasedandcellviabilitywasdecreasedinculturedastrocytetreatedbyH／RIncontrasttocontrolcells(H／Rgroups)，thrombinsignificantlyenhancetheeffectsofH／Rinadosedependentmanner(P005)③IncubationofH瓜astrocyte

6、withTB(10U／m1)，enhancedthee艉吐ofH／RstimulationComparingwithcontrolcellsandtheenhancementofthrombinincreaseswiththetimeofreoxygenation④InjuryofH／RinducedthereleaseofNOthedecreaseofcellsviabilityandapoptosis，theseeffectsWas

7、aggravatedbythrombinHirudincallreversetheeffectofthrombin(9ImPCRandimmunocytochemistryanalysisdemonstratedthattheIevelofiNOSmRNAandiNOSproteinincreasedinastrocytetreatedbyH／RTBenhancedtheexpressionofiNOSmRNAandiNOSprotei