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1、rhorock信号通路介导缺氧所致人肺微血管内皮细胞肌动蛋白细胞骨架重构和重组人白介素10对缺氧条件下人肺微血管内皮细胞凋亡改变规律的影响广州医学院硕士学位论文5Hypoxia-InducedReorganizationofActinCytoskeletonMediatedbyRho-ROCKSignalPathwayinHumanPulmonaryMicrovascularEndothelialCellsandTheEffectofRecombinationHumanInterleukin10onHypoxia-inducedApoptosisinHumanPu1nonaryMicrovas
2、cularEndothelialCellsProf.Qi-IiangCuiCandidate:JingMoSupervisor:Abstract1Objective1.1ToinvestigatetheroleofRho-ROCKsignalpathwayinthereorganizationofactincytoskeletonandthechangesofmorphologyinducedbyhypoxiainhumanpulmonarymicrovascularendothelialcelIs(HPMVECs).1.2Toinvestigatetheameliorativemechani
3、smofrecombinatinghumaninterleukin10(rhI1.-10)throughinhibitingtheapoptosisandCaSPaSe-3activityinhypoxia-inducedhumanpulmonarymicrovascularendothelialcelIs.2MaterialsandMethods2.1Duringtheexperimentsthehumanpulmonarymicrovascularendothelialcellswereexposedtohypoxiafor6h,12hand24hrespectivelyorpre-inc
4、ubatedwithaspecialinhibitorofKOCK,H-1152(10nmol1.)for0.5hbeforehypoxiainvitro.ThereorganizationofcellularactincytoskeletonandthechangesofcellularmorphologyinducedbyhypoxiaorpretreatmentofHl152wereexaminedbylaserscanningconfocal广州医学院硕士学位论文6microscopy(1.SCM).TheF-actinmeanfluorescenceintensityofeachgr
5、oupwasexaminedbyflowcytometric.2.2TheculturedHPMVECswererandomizedtonormalcontrolgroup,hypoxiacontrolgroup,thelow-rhI1.-10interventiongroup,middle-rhI1.-10interventiongroupandhigh-doserhI1.-10interventiongroup.Thenormalgroupwasculturedwiththe37eCand5%CO2condition,andthehypoxiagroupswereculturedwith5%CO2,95%N2condition.Theculturedtimelastedfor4h,12h,24hand48hrespectively.Therelativecaspase-3activityofal1groupsweredetectedwithChromatometryattheendof4h,12h,24hand48h.Theapoptosisfluorescenceintensitywereanalyzedbythefluorescentmicroscopeattheendof24hwithstatisticalmet.