Effect of Testosterone on Insulin Stimulated IRS1 Ser Phosphorylation in Primary Rat Myotubes—A Potential Model for PCOS-Related Insulin Resistance 英文参考文献.docVIP

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Effect of Testosterone on Insulin Stimulated IRS1 Ser Phosphorylation in Primary Rat Myotubes—A Potential Model for PCOS-Related Insulin Resistance 英文参考文献.doc

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Effect of Testosterone on Insulin Stimulated IRS1 Ser Phosphorylation in Primary Rat Myotubes—A Potential Model for PCOS-Related Insulin Resistance 英文参考文献

EffectofTestosteroneonInsulinStimulatedIRS1Ser PhosphorylationinPrimaryRatMyotubes—APotential ModelforPCOS-RelatedInsulinResistance MichaelC.Allemand1,BrianA.Irving2,YanW.Asmann2,KatherineA.Klaus2,LauraTatpati1,CharlesC. Coddington1,K.SreekumaranNair2* 1DepartmentofObstetricsandGynecology,DivisionofReproductiveEndocrinologyandInfertility,MayoClinicGraduateSchoolofMedicine,Rochester,Minnesota,United StatesofAmerica,2DepartmentofInternalMedicine,EndocrineResearchUnit,MayoClinicGraduateSchoolofMedicine,Rochester,Minnesota,UnitedStatesofAmerica Abstract Background: Polycystic ovary syndrome (PCOS) is characterized by a hyperandrogenic state and frequently develops skeletalmuscleinsulinresistance.Wedeterminedwhethertestosteroneadverselyaffectsinsulinactionbyincreasingserine phosphorylationofIRS-1636/639 indifferentiatedratskeletalmusclemyotubes.ThephosphorylationofAkt,mTOR,andS6K, downstreamtargetsofthePI3-kinase-IRS-1complexwerealsostudied. Methods:Primarydifferentiatedratskeletalmusclemyotubesweresubjectedtoinsulinfor30minafter16-hourpre-exposure toeitherlow(20ng/ml)orhigh(200ng/ml)dosesoftestosterone.ProteinphosphorylationofIRS-1Ser636/639,AktSer473 , mTOR-Ser2448,andS6K-Thr389weremeasuredbyWesternblotwithsignalintensitymeasuredbyimmunofluorescence. Results:Cellsexposedto100nMofinsulinhadincreasedIRS-1Ser636/639andAktSer473phosphorylation.Cellspre-exposed to low-dose testosterone had significantly increased insulin-induced mTOR-Ser2448 and S6K-Thr389 (p,0.05),andfurtherincreasedinsulin-inducedIRS-1Ser636/639phosphorylation(p=0.042)comparedtocontrolcells.High- dosetestosteronepre-exposureattenuatedtheinsulin-inducedmTOR-Ser2448 andS6K-Thr389 phosphorylation phosphorylation. Conclusions:Thedatademonstratedaninteractionbetweentestosteroneandinsulinonphosphorylationofintracellular signalingproteins,andsuggestsalinkbetweenahyperandrogenic,hyperinsulinemicenvironmentandthedevelopmentof insulin resistance involving serine phosphorylation of IRS-1 Ser636/639. These resul