实验性氟中毒中枢神经系统炎症损伤机制中nf-b信号通路改变的作用-role of nf - b signaling pathway changes in inflammatory injury mechanism of central nervous system in experimental fluorosis.docxVIP

显示，用500及5000?mol/LNaF培养细胞24、48及72小时后，500?mol/LNaF及以上浓度可明显降低THP-1细胞存活率，呈现存活率与时间和剂量的负相关关系；细胞凋亡率随着染氟时间的延长逐渐增高；低、高剂量染氟组THP-1细胞炎症因子TNF?和IL-1?含量明显高于对照组（P0.05）；NF-κBp65、IκBα总蛋白表达均明显低于对照组（P0.05），而Phospho-NF-κBp65和Phospho-IκB-α蛋白表达均明显高于对照组（P0.05）；相关分析结果显示，低、高剂量染氟组THP-1细胞中Phospho-NF-κBp65表达水平与TNF?、IL-1?含量成正比，与IκBα表达成反比。结论：慢性氟中毒大鼠脑组织及过量氟暴露的体外培养细胞可产生大量的炎性因子及NF-κB信号通路激活，可能是过量的氟化钠蓄积导致中枢神经系统损伤的机制之一。【关键词】：慢性氟中毒；大鼠；胶质细胞；NF-?B通路；TNF?IIIEffectofchangedNF-?BsignalpathwayinmechanismofinflammationinjuryinthecentralnervoussysteminducedbyexperimentalfluorosisMajor:PathologyandPathophysiologyPostgraduate:TANGTing-tingSupervisor:GUANZhi-zhong[Abstract]Objective:ToinvestigatetheexpressionofnuclearfactorkappaBsignaltransductionpathwayofNF-kappaBp65,nuclearfactorkappaBinhibitorprotein,Phospho-NF-kappaBp65andPhospho-IkappaB-alphainchronicfluorosisanimalandcellmodels.Toexplorethepossiblepathologicalmechanismofinflammatoryinbraindamageofratsduringchronicfluorineandprovideatheoreticalbasisforthepreventionofendemicfluorosis.Methods:Establishtheanimalmodelofchronicfluorosis:SixetySDratsweredividedrandomlytothreegroupsof30each(ahalffemalesandahalfmales),e.g.thecontrolgroupfedwithnormaltap-water,high-fluoridegroupwith50mg/Lfluorideaddedindrinkingwater.Theexperimentperiodwas10months.Theestablishthecellmodelofchronicfluorosis:Theexperimentsweredividedintofivegroups,seedTHP-1cellsin6poreplate,e.g.thecontrolgroup,thelowgroupwith500?mol/LNaF,thehighgroupwith5000?mol/L.ThefluoridecontentsinurineandboneweredetectedbyFluoride-ionselectiveelectrode.TestingtheratslearningandmemoryfunctionbyMorriswatermazeexperiment.Observationofbraintissuestructureunderlightmicroscope;TheexpressionofCD11bandGFAPbyimmunohistochemistry.Concentrationsofinflammatorycytokines,suchasIL-1βandTNFαweremeasuredbyEnzymelinkedimmunosorbentassay(Elisa)inbraintissue.TheproteinlevelsofNF-κBp65,IκBα,Phospho-NF-κBp65andPhospho-IκB-αweredetectedbyWesternBlotting.CelltoxicitytestbyCCK8.TheapoptosisofTHP-1cellswerebyflowcytomety.Results:1.Theresult