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Notch通路对高糖诱导人晶状体上皮细胞凋亡中P53、BAX作用的研究.docx

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Notch通路对高糖诱导人晶状体上皮细胞凋亡中P53、BAX作用的研究 Title:RoleofNotchSignalingPathwayinHighGlucoseInducedApoptosisofHumanLensEpithelialCells:FocusonP53andBAX Abstract: Diabeticcataractisacommoncomplicationofdiabetesmellitus,characterizedbyprogressiveopacificationofthelens.Theunderlyingmechanismsofdiabeticcataractremainunclear.ThisstudyaimedtoinvestigatetheroleoftheNotchsignalingpathwayinhighglucose-inducedapoptosisofhumanlensepithelialcells(HLECs),focusingontheinvolvementofP53andBAXproteins. Introduction: Diabeticcataractisamajorcauseofvisualimpairment,oftenoccurringinpatientswithpoorlycontrolleddiabetes.Highglucoselevelshavebeenimplicatedasamajorcontributortothedevelopmentofdiabeticcataract,leadingtooxidativestressandcelldeathinlensepithelialcells.Notchsignalinghasemergedasacriticalpathwayregulatingcellfatedecisionsandapoptosisinvariouscellularcontexts.However,itsroleinhighglucose-inducedapoptosisinHLECsremainslargelyunexplored.ThisstudyaimstobridgethisknowledgegapbyinvestigatingtheinvolvementofP53andBAXproteinsinNotchsignaling-mediatedapoptosisinHLECsunderhighglucoseconditions. Methods: 1.Cellculture:Humanlensepithelialcells(HLECs)willbeculturedanddividedintocontrolandhighglucosegroups. 2.Highglucosetreatment:HLECsinthehighglucosegroupwillbetreatedwithahighglucosemediumtomimicthediabeticcondition. 3.Notchsignalingmodulation:HLECsinthehighglucosegroupwillbetreatedwithNotchsignalinginhibitorsoractivatorstoassesstheimpactonapoptosis. 4.Detectionofapoptosis:ApoptosiswillbeassessedusingAnnexinV/propidiumiodidestainingandflowcytometry. 5.Quantitativereal-timepolymerasechainreaction(qRT-PCR):TheexpressionlevelsofgenesrelatedtoNotchsignaling,P53,andBAXwillbedetermined. 6.Westernblotanalysis:ProteinexpressionlevelsofNotchsignalingcomponents,P53,andBAXwillbeevaluated. 7.Statisticalanalysis:Datawillbeanalyzedusingappropriatestatisticalmethodstodeterminesignificantdifferencesbetweengroups. Results: OurpreliminaryfindingsdemonstratedthathighglucosetreatmentincreasedHLECapoptosiscomparedtothec