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1、XBP1對脂多糖處理的Kupffer細(xì)胞炎性因子表達的影響李鐵軍1，劉作金2，李生偉2，劉長安2，屈謙3，薛強2，龔建平2（400700重慶，1重慶市第九人民醫(yī)院普外科三科；400010重慶，2重慶醫(yī)科大學(xué)附屬第二醫(yī)院肝膽外科市；401147重慶，重慶市衛(wèi)生局）[摘要]目的探討XBP1對脂多糖(LPS)處理的Kupffer細(xì)胞(kupffercells，KCs)TNF-α表達的影響。方法將BALB/c小鼠KCs隨機分為質(zhì)粒轉(zhuǎn)染組（質(zhì)粒組），陰性對照組（陰性組）和空白對照組（空白組）。分別給予XBP1-s
2、hRNA，空白質(zhì)粒轉(zhuǎn)染和空白處理，并給予脂多糖。免疫細(xì)胞化學(xué)（SABC法）、Realtime-PCR和Western-blot法檢測各組KCs中XBP1基因及蛋白表達水平；酶聯(lián)免疫吸附法（ELISA）檢測核因子NF-κB活性及培養(yǎng)上清液中TNF-α含量。結(jié)果與兩對照組相比，質(zhì)粒組XBP1及下游因子TNF-α表達明顯降低（P<0.05）；而兩對照組間無明顯差異（P>0.05）。3組間NF-κB活性無明顯差異（P>0.05）。結(jié)論XBP1-shRNA質(zhì)粒能通路明顯抑制KCsXBP1基因的表達并通過非NF-κ
3、B通路減少其下游因子TNF-α的釋放。[關(guān)鍵詞]X盒結(jié)合蛋白l；Kupffer細(xì)胞；腫瘤壞死因子-α；小鼠[中圖發(fā)分類號][文獻標(biāo)志碼]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；
