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屎肠球菌HDRsEf1调节IPEC-J2细胞炎症反应机制研究的开题报告.docx

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屎肠球菌HDRsEf1调节IPEC-J2细胞炎症反应机制研究的开题报告 Title：StudyontheMechanismofStreptococcussuisHDRsEf1RegulatingInflammatoryResponseinIPEC-J2Cells Introduction Streptococcussuis(S.suis)isazoonoticpathogenthatcancausesevereinfectionsinpigsandhumans.Inrecentyears,theprevalenceofS.suisinfectionshasincreasedworldwide,leadingtoeconomiclossesandpublichealthconcerns.InflammatoryresponseplaysanimportantroleinthepathogenesisofS.suisinfection.Intestinalepithelialcells(IECs)serveasacrucialfirstlineofdefenseagainstpathogenicbacteriaandareinvolvedininitiatingandregulatinginflammatoryresponses.IPEC-J2cellline,anon-transformedIECmodelderivedfrompigjejunum,hasbeenwidelyusedtoinvestigatethemechanismsofIEC-mediatedimmunityandinflammation. HypotheticalModel HDRsEf1,aspecifichistidinekinaseresponseregulatorinS.suis,hasbeenreportedtobeassociatedwiththevirulenceofthisbacterium.ItishypothesizedthatS.suisHDRsEf1mayregulatetheinflammatoryresponseinIPEC-J2cellsthroughasignaltransductionpathwaymediatedbytheTLR4/NF-κBsignalingpathway. Objectives 1.ToevaluatetheeffectofS.suisHDRsEf1ontheproductionofinflammatorycytokinesinIPEC-J2cells. 2.ToinvestigatethesignalingpathwayinvolvedintheregulationofinflammatoryresponsebyS.suisHDRsEf1inIPEC-J2cells. Methods 1.IPEC-J2cellswillbestimulatedwithS.suisHDRsEf1andtheexpressionlevelsofIL-1β,IL-6,andTNF-αwillbemeasuredbyRT-qPCRandELISA. 2.IPEC-J2cellswillbetreatedwithTLR4inhibitorandNF-κBinhibitorfollowedbystimulationwithS.suisHDRsEf1,andtheexpressionlevelsofIL-1β,IL-6,andTNF-αwillbemeasured. 3.WesternblottingwillbeusedtoinvestigatetheactivationoftheTLR4/NF-κBsignalingpathwayinIPEC-J2cellsstimulatedwithS.suisHDRsEf1. ExpectedResults ItisexpectedthatS.suisHDRsEf1willinducetheproductionofinflammatorycytokinesinIPEC-J2cells.ByblockingtheTLR4/NF-κBsignalingpathwaywithinhibitors,itisexpectedthattheproductionofinflammatorycytokineswillbereduced.ItisalsoexpectedthatS.suisHDRsEf1willactivatetheTLR4/NF-κBsignalingpathwayinIPEC-J2cells. Conclusion ThisstudyaimstoexplorethemechanismofS