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时间:2020-05-15
《芹菜素抑制脂多糖诱导小鼠巨噬细胞分泌炎症介质的分子机制.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、吴广等芹菜素抑制脂多糖诱导小鼠巨噬细胞分泌炎症介质的分子机制第6期·753·doi:10.3969/j.issn.1000-484X.2015.06.007芹菜素抑制脂多糖诱导小鼠巨噬细胞分泌炎症介质的分子机制①吴广符平②周玉生周润梅③(南华大学附属第二医院药剂科,衡阳421001)中图分类号R967文献标志码A文章编号1000-484X(2015)06-0753-05[摘要]目的:观察芹菜素对脂多糖(LPS)诱导小鼠巨噬细胞分泌炎症介质的影响,并探讨其分子机制。方法:体外培养小鼠巨噬细胞RAW264.7,用不同浓度的芹菜素处理后,加入LPS刺激。Westernb
2、lot检测血红素氧合酶.1(HO.1)、环氧化酶-2(COX一2)、诱导型一氧化氮合成酶(iNOS)的表达以及p38和IKB的磷酸化;比色法检测亚硝酸盐和硝酸盐(NOx)的含量;ELISA检测PGE2的产生;报告基因分析芹菜素对NF-KB激活的影响。结果:芹菜素可显著诱导RAW264.7细胞表达HO.1,采用p38抑制剂SB203580预处理细胞后,可抑制芹菜素诱导的HO一1表达;同时,芹菜素能降低胞浆内核转录因子Nrf2含量,并增高其在细胞核内水平;采用siRNA干扰Nri2表达后,HO·1表达量降低。此外,芹菜素也能抑制LPS诱导RAW264.7细胞产生NOx
3、和PGE2,下调COX-2和iNOS表达,以及抑制IKB磷酸化和NF-KB激活。而用HO一1siRNA转染后,能在一定程度上抑制芹菜素的上述效应。结论:芹菜素可能通过诱导HO一1表达、抑制NF-KB的激活而发挥抗炎症作用。[关键词]芹菜素;血红素氧合酶一1;小鼠巨噬细胞MolecularmechanismofapigeninoninhibitionofLPS-inducedinflammatorymediatorsinmurinemacrophagesWUGuang。FUP讯g,zHoUYu—Sheng,ZHOURun—Mei.DepartmentofPharma
4、cy,theSecondAffiliatedHospital,UniversityofSouthChina,Hengyang421001,China[Abstract]Objective:ToinvestigatetheeffectandthemechanismofApigeninonliPopolysaccharides(LPS)一inducedinflammatorymediatorsproductioninmurinemacrophages.Methods:ThemurinemacrophagecelllineRAW264.7cellswereculture
5、dinvitro,andweretreatedwithdiferentconcentrationofApigeninfollowedbyLPSadministration.Expressionofhemeoxygenase一1(HO一1),cyclooxygenase-2(COX-2)andinduciblenitricoxidesynthase(iNOS),phosphorylationofp38andIKB,nucleartranslocationofNrf2weredetectedbyWesternblot.ProductionofNitriteandnit
6、rate(NOx)wasanalyzedbycolorimetrietechnique.Secretionofprosta—glandinE2(PGE2)wasdetectedbyELISA.ActivationofNF-KBwasmeasuredbyluciferaseassay.Results:WesternblotindicatedthatapigenincouldinduceRAW264.7cellsexpressionofHO一1,andpretreatmentofSB203580,aninhibitorofp38significantlyinhibit
7、edapigenininducedHO-1expression.Inaddition,ApigenincouldalsodecreasethecontentofnucleartranscriptionfactorNrf2incytoplasmandincreaseitslevelinthenucleus.SilencingofNrf2byspecificsiRNAcouldinhibitapigenin·inducedHO一1expression.Furthermore,apigeninadministrationsignificantlyinhibitedLPS
8、·indu
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