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1、XBP1对脂多糖处理的Kupffer细胞炎性因子表达的影响李铁军1，刘作金2，李生伟2，刘长安2，屈谦3，薛强2，龚建平2（400700重庆，1重庆市第九人民医院普外科三科；400010重庆，2重庆医科大学附属第二医院肝胆外科市；401147重庆，重庆市卫生局）[摘要]目的探讨XBP1对脂多糖(LPS)处理的Kupffer细胞(kupffercells，KCs)TNF-α表达的影响。方法将BALB/c小鼠KCs随机分为质粒转染组（质粒组），阴性对照组（阴性组）和空白对照组（空白组）。分别给予XBP1-s
2、hRNA，空白质粒转染和空白处理，并给予脂多糖。免疫细胞化学（SABC法）、Realtime-PCR和Western-blot法检测各组KCs中XBP1基因及蛋白表达水平；酶联免疫吸附法（ELISA）检测核因子NF-κB活性及培养上清液中TNF-α含量。结果与两对照组相比，质粒组XBP1及下游因子TNF-α表达明显降低（P<0.05）；而两对照组间无明显差异（P>0.05）。3组间NF-κB活性无明显差异（P>0.05）。结论XBP1-shRNA质粒能通路明显抑制KCsXBP1基因的表达并通过非NF-κ
3、B通路减少其下游因子TNF-α的释放。[关键词]X盒结合蛋白l；Kupffer细胞；肿瘤坏死因子-α；小鼠[中图发分类号][文献标志码]ATheinfluenceofXBP1onexpressionofinflammatoryfactorofLPS-inducedKupffercellsLiTiejun1，LiuZuojin2，LiShengwei2，LiuChang’an2，QuQian3，XueQiang2，GongJianping2(DepartmentofGeneralSurgery,ninth
4、People'sHospital,Chongqing1,4007001;DepartmentofHepatobiliarySurgery,theSecondAffiliatedHospital,ChongqingMedicalUniversity,Chongqing2,400010;HealthBureauofChongqing4,401147,China)[Abstract]ObjectiveToexploretheinfluenceofXBP1onexpressionofinflammatoryfa
5、ctorofLPS-inducedKupffercells.MethodsKCsderivedfromBALB/cmicewererandomlydividedintoXBP1-shRNAgroup,negativecontrolgroup(negativegroup)andblankcontrolgroup(blankgroup),whichweretreatedwithXBP1-shRNAplasmidtransfection,controlvectortransfectionandnotreatm
6、ent,respectively,andthentreatedwithLPS.ThemRNAandproteinexpressionofXBP1inKCsweredeterminedbyreal-timePCR,western-blotandimmunohistochemistry(SABC).ThedensityofTNF-αinthesupernatantofcocultureandNF-κBwasassessedbyenzyme-linkedimmunosorbentassay(ELISA).Re
7、sultsTheexpressionofXBP1anditsdownstreamfactorTNF-αweresignificantlylowerthanthoseinnegativegroupandblankgroup(P<0.05).However,Therewerenosignificantdifferencebetweentwocontrolgroups(P>0.05).Inaddition,theactivitiesofNF-κBinthreegroupshadnosignificantdif
8、ference(P>0.05).ConclusionXBP1-shRNAplasmidcansignificantlyinhibittheexpressionofXBP1geneandreducethereleaseofdownstreamfactorTNF-αwithoutchangingtheactivitiesofNF-κBinKCs.[keywords]X-boxbindingprotein1；Kupffercells；TNF-α；
