AMPK通過轉(zhuǎn)錄因子SP1影響下丘腦GT1-7神經(jīng)元KiSS-1基因轉(zhuǎn)錄的機制研究.pdf

1、福建醫(yī)科大學碩士學位論文AMPK通過轉(zhuǎn)錄因子SP1影響下丘腦GT17神經(jīng)元KiSS1基因轉(zhuǎn)錄的機制研究姓名：劉春娥申請學位級別：碩士專業(yè)：內(nèi)科學(內(nèi)分泌與代謝)指導教師：陳剛；溫俊平2011036AMPKinhibitsKiSS1genetranionexpressionthroughtranionalfactSP1inthehypothalamicGT17neuronsAbstractObjective:Thestudywastoin

2、vestigatetheeffectoftranionalfactSP1onthepromoteractivityofKiSS1geneinthehypothalamusGT17neuronsaswellastoexpletheroleofAMPKplayedinthetranslocationofSP1inthesecells.Methods:(1).WetransfectedGT17cellswiththe2kbfulllength

3、KiSS1genepromoter(PGL3KiSS1promoter)linkedwithaluciferaserepterthentreatedthemwithSP1SiRNAMithramycinAknownasapharmacologicalsuppressofSP1detectedtheluciferaseactivitybyDualLuciferaseRepterAssaySystem.(2).WeincubatedGT17

4、cellswithAICAR(anartificialactivatofAMPK)CompoundC(asuppressofAMPK)AMPKα1siRNAdeterminedthelevelsofSP1proteinfromnuclearfractioncytoplasmicfractiontotalcellextractsbyWesternBlot.(3).WetreatedGT17cellswithAICARCompoundCpe

5、rfmedtheimmunofluescenceanalysistodirectlyvisualizetheeffectofAMPKonthesubcellularredistributionofSP1protein.Results:(1).SP1SiRNAMithramycinAatdifferentconcentrationscoulddecreasethepromoteractivityofKiSS1geneobviouslyin

6、thehypothalamusGT17cells.(2).AICARdecreasedthenuclearfractionincreasedthecytoplasmfractionofSP1.TothecontraryCompoundCAMPKα1SiRNAplayedaninverserolecomparedwithAICARinthesubcellularcompartmentalizationofSP1inGT17cells.AI

7、CARCompoundCaswellasAMPKα1SiRNAhadnosignificanteffectonthetotalextractsofSP1intheGT17cells.(3).TheresultsfromourimmunofluescenceanalysisshowedthatAICARinhibitedthetranslocationofSP1whileCompoundCenhancedthetranslocationo