2的表达及AP-1、NFKB信号通路.doc

2的表达及AP-1、NFKB信号通路 中国医学科学院 ACIlAACADEMIAEMEDICINAESINICAE 致密斑细胞COX-2的表达及AP-1,NFKB信号通路 刘冬妍,李学旺,李航,李雪梅,叶文玲 中国医学科学院中国协和医科大学北京协和医院肾内科,北京100730 通信作者:李学旺电子邮件:leexuewang@hotmail.corn ? 论着? 摘要:目的评估低盐(LS)培养对小鼠致密斑(MMDDI)细胞环氧化酶一2(cOx-2)表达及核因子-KB (NF.KB)和活化蛋白.1(AP_1)活性的影响.方法经脂质体转染含NF-KB或AP-I的报告质粒,采用瞬时表达方法检 测正常盐(NS)与LS培养对NF.KB和AP-l转录活性的影响.采用RT-PCR检测MMDDI细胞C0x-2表达的变化, Westernblot方法检测细胞内p.p38MAPK,p-p44/42,c-Jun,c.Fos和C0x-2蛋白的表达.结果LS培养促进了MMDDI 细胞c0x-2mRNA和蛋白表达(Plt;0.01).LS培养后,p38和p44/42的磷酸化程度显着上调(Plt;0.01),180rain后 达到高峰.p38抑制剂SB-203580,p44/42抑制剂PD-98059可降低LS诱导的C0x-2表达(Plt;0.01).LS培养促进了 c.Jan,c.Fos蛋白表达(Plt;0.叭),激活了AP?l和NF-KB的转录活性(Plt;0.叭).25~moVLNF-KB抑制剂PDTC和20 p,mol/LAP.1抑制剂curcumin下调了LS诱导的NF-KB,AP-I活性(Plt;0.叭).25p,mol/LPDTC,20p,mol/Lcurcumin 降低了LS诱导的c0x-2mRNA和蛋白表达(Plt;0.01).结论LS培养可促进MMDDI细胞c0x-2的表达,其作用可 能与促进p38MAPK,p44/42激酶的磷酸化,增加NF.KB和AP_l的活性有关. 关键词:环氧化酶一2;核因子-KB;活化蛋白.1;质粒;丝裂素激活蛋白激酶 中围分类号:R334.1文献标识码:A文章编号:1000-503X(2007)01-0078-05 ExpressionofCyciooxygenase~inaMouseMaculaDensaCell LinesandSignalTransductionofNF?-KBandAP?-1 LIUDong—yan,LIXue—wang,LIHang,LIXue—mei,YEWeng—ling DepartmentofNephrology,PUMCHospital,CAMSandPUMC,Beijing100730,China Correspondingauthor:UXue—wangE—mail:leexuewang@hotmail.corn ABSTRACT:ObjectiveToelvaluatetheeffectoflowsalt(LS)ontheexpressionofcyclooxygenase-2 (COX-2)andtheactivityofnuclearfactorkappaB(NF—Ks)andactivatorprotein一1(AP一1)inthemouse maculadensaderived(MMDDI)cellline.MethodsMMDDIcellsweretransfectedwithluciferasereporter plasmidcontainingAP一1orNF—KB.Luciferasereporterassaywasusedtoevalu~etheeffectofnormalsalt (NS)andlOWsalt(LS)ontheactivitiesofNF.KBandAP.1.ThechangesofCOX-2expressionwereexam. inedbyRT—PCR.Theexpressionofp-p38MAPK,p-p44/42,c—Jun,c—Fos,andCOX-2inMMDDIcells wereanalyzedbyWesternblot.ResultsTheexpressionsofCOX一2mRNAandproteininMMDD1cellswere significantlyincreasedbyLS(Plt;0.O1).Phosphorylatedp38andp44/42MAPkinaseweresignificantlyin— creasedbytreatmentat180min(Plt;0.01).Theup—regulatedCOX-2proteinexpressionwithLSweresignifi— cantlyr