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1、Trim22siRNA對LPS誘導的巨噬細胞促炎細胞因子的影響孫大康1,安新業(yè)2,趙延婷2,宋向芹2(256603山東濱州,濱州醫(yī)學院附屬醫(yī)院:臨床醫(yī)學實驗中心1,檢驗科2)[摘要]目的探討Trim30?旁系同源分子Trim22特異性siRNA對脂多糖(LPS)誘導的巨噬細胞促炎細胞因子TNF?和IL6表達的影響。方法干擾素?(IFN?)刺激人單核細胞白血病細胞株U937,觀察Trim22mRNA表達水平的變化;用佛波酯(PMA)將U937細胞誘導分化為巨噬細胞,觀察LPS刺激巨噬細胞時IL6表達的時相變化。用靶向Trim22的si
2、RNA預處理,ELISA觀察對LPS誘導的巨噬細胞促炎細胞因子TNF?和IL6表達的影響。結(jié)果IFN?可以顯著促進U937細胞Trim22的mRNA表達,同時U937細胞誘導分化的巨噬細胞受LPS刺激時IL6顯著升高。靶向Trim22的siRNA可以顯著提高LPS誘導的巨噬細胞TNF?和IL6的表達。結(jié)論Trim22對LPS誘導的巨噬細胞促炎細胞因子TNF?和IL6具有負向調(diào)節(jié)作用。[關(guān)鍵詞]Trim22;RNA干擾;脂多糖;巨噬細胞;促炎細胞因子[中圖分類號]R392.11[文獻標志碼]AEffectofTrim22siRNAon
3、LPS-mediatedproinflammatorycytokinesexpressioninmacrophagesSUNDakang1,ANXinye2,ZHAOYanting2,SONGXiangqin2(1ExperimentCenterofClinicalMedicine,2ClinicalLaboratory,AffiliatedHospitalofBinZhouMedicalUniversity,BinZhou256603)[Abstract]0bjectiveToinvestigatetheeffectofsiRNA
4、targetingTrim22,oneoftheparaloguesofTrim30?,onLPS-mediatedproinflammatorycytokinesexpressioninmacrophages.MethodsTheexpressionofTrim22inducedbyIFN?wasexaminedbyRT-PCRinhumanpromonocyticU937cells.Then,U937cellsweredifferentiatedintomacrophagesbyPMAandLPS-inducedproinfla
5、mmatorycytokineIL6levelsweremeasuredbyELISA.MacrophagesweretransfectedwithsiRNAtargetingTrim22;subsequentlycellswerestimulatedwith500U/mlofIFN?for24handchallengedwithLPStodetermineproinflammatorycytokinesexpression.ResultsExpressionofTrim22showedanincreaseinU937cellsac
6、tivatedwithIFN?.Moreover,LPSstimulationcausedpronouncedupregulationofIL6inMacrophages.Asaconsequence,suppressionofTrim22activitybysmallinterferingRNAenhancesTNF?,IL6productioninMacrophages.ConclusionTrim22negativeregulateLPS-mediatedproinflammatorycytokinesexpressionin
7、macrophages.[Keywords]Trim22,siRNA;Lipopolysaccharide;macrophages;proinflammatorycytokinesSupportedbyTheProjectofShandongProvinceHigherEducationalScienceandTechnologyProgram(J11LF89),ProgramofScienceandTechnologyDevelopmentofYan'tai(2011078).Correspondingauthor:SunDaka
8、ng,Tel:0543-3258709,E-mail:sdkaaa@163.com目前人類已知有70多種三基序蛋白(tripartite-motifprotein,Trim)家族成員,具有重要的抗病毒[1]作用,并參與細胞的增值、分化