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1、內(nèi)毒素對(duì)離體鼻黏膜上皮細(xì)胞IFNγ、IL4表達(dá)的影響【摘要】目的:探討內(nèi)毒素(LPS)對(duì)鼻黏膜上皮細(xì)胞(HNEC)IFNγ、IL4表達(dá)的影響。方法:設(shè)LPS組、LPS+地塞米松(DXM)組、LPS+吡咯啉烷二硫代氨基甲酸鹽(PDTC)組及生理鹽水空白對(duì)照組。采用酶聯(lián)免疫吸附及原位雜交方法檢測(cè)培養(yǎng)上清液中IFNγ、IL4及HNEC內(nèi)NFкBp50mRNA的表達(dá),凝膠電泳遷移率檢測(cè)NFкBp50,觀察PDTC及DXM對(duì)NFкBp50活化抑制后IFNγ和IL4蛋白水平。結(jié)果:(1)0~100μg/LLPS隨濃度的增加,IFNγ分泌
2、量增多;>100μg/L后,IFNγ分泌量減少;100μg/LLPS刺激HNEC分泌IFNγ的作用最強(qiáng)。0~20μg/LLPS隨濃度的增加,IL4分泌量增多;>20μg/L后,IFNγ分泌量減少;20μg/LLPS刺激HNEC分泌IL4的作用最強(qiáng),40μg/LLPS刺激IFNγ和IL4的分泌有交叉,二者維持于一定的水平。(2)50μg/LLPS刺激HNEC后隨時(shí)間延長(zhǎng)IFNγ分泌量逐漸增加,從4h開(kāi)始即有明顯上升(P<0.05),8h達(dá)到峰值;IL4隨時(shí)間延長(zhǎng)分泌量逐漸增加,從2h開(kāi)始即有明顯上升(P<
3、0.05),6h達(dá)到峰值。(3)PDTC和DXM均抑制NFкBp50的表達(dá)和活性,減少HNEC分泌IFNγ和IL4,DXM4.0μg/L時(shí),下降更明顯,但高于此濃度時(shí),不引起進(jìn)一步的降低。結(jié)論:12LPS對(duì)HNEC的刺激反應(yīng)與濃度呈劑量依賴性,適當(dāng)濃度的LPS刺激有利于HNEC分泌的Th1/Th2細(xì)胞因子維持平衡狀態(tài);過(guò)高或過(guò)低濃度的LPS刺激可誘導(dǎo)HNEC的Th1/Th2細(xì)胞因子失衡分泌,這可能是GNB在慢性鼻、鼻竇炎(CRS)的致病機(jī)理?!娟P(guān)鍵詞】?jī)?nèi)毒素類;上皮細(xì)胞;鼻黏膜;核因子кBp50;IFNγ;白細(xì)胞介素4 ?。跘bstra
4、ct]Objective:Tostudytheinfluenceofendoutoxin(lipopolysaccharide,LPS)toexpressionofIFNγandIL4inhumannasalepithelialcell(HNEC).Methods:CulturedHNECweredividedintogroupLPS,groupDXM,andgroupPDTC,thatwereaddedwithLPS,LPSplusdexamethasone,andLPSpluspyrrolinedithiocarbamaterespec
5、tively.Ablankcontrolgroup(groupB)wassetupbyaddingnormalsaline.ExpressionofIFNγandIL4insupernatantsofculturemediaandNFкBp50mRNAexpressioninHNECweredetectedwithenzymelinkedimmuneadsorption(ELISA)andhybridizationinsitu;TheactivationofNFкBp50wasdetectedbygelelectrophoresism
6、obilityanalysis;TheproteinlevelchangesofIFNγandIL4aftertheinhibitionofPDTCandDXMtoNFкBp50wasobserved.Result:LPScoulddirectlyactontheHNECtopromotethesecretionsofIFNγandIL4intermsoftime.TheLPSexcitementcouldsignificantlypromotetheactivationofNFкBp50,whichpeakedin3hoursan
7、dthenwentdown.PDTCandDXM12couldsignificantlydecreaseanalogicallythesecretionsofIFNγandIL4andtheactivationofNFкBp50.Conclusions:TheresponseofHNECtothestimulationofLPSshowsdosagedependence,andstimulationofLPSwithsuitableconcentrationbenefitsHNECsecretingTh1/Th2cellfactorsto
8、maintainthebalance;ThestimulationofLPSwithtoohighandtoolowconcentrationswou