Electrical Pulse Stimulation of Cultured Human Skeletal Muscle Cells as an In Vitro Model of Exercise 英文参考文献.docVIP

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Electrical Pulse Stimulation of Cultured Human Skeletal Muscle Cells as an In Vitro Model of Exercise 英文参考文献.doc

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Electrical Pulse Stimulation of Cultured Human Skeletal Muscle Cells as an In Vitro Model of Exercise 英文参考文献

ElectricalPulseStimulationofCulturedHumanSkeletal MuscleCellsasanInVitroModelofExercise Natasˇa Nikolic′1*,SirilSkaretBakke1,EiliTranheimKase1,IdaRudberg1,2,IngeborgFloHalle1,ArildC. Rustan1,G.HegeThoresen1,VigdisAas2 1DepartmentofPharmaceuticalBiosciences,SchoolofPharmacy,UniversityofOslo,Oslo,Norway,2FacultyofHealthSciences,OsloandAkershusUniversityCollegeof AppliedSciences,Oslo,Norway Abstract BackgroundandAims:Physicalexerciseleadstosubstantialadaptiveresponsesinskeletalmusclesandplaysacentralrole inahealthylifestyle.Sinceexerciseinducesmajorsystemicresponses,underlyingcellularmechanismsaredifficulttostudy invivo.Itwasthereforedesirabletodevelopaninvitromodelthatwouldresembletraininginculturedhumanmyotubes. Methods: Electrical pulse stimulation (EPS) was applied to adherent human myotubes. Cellular contents of ATP, phosphocreatine(PCr)andlactateweredetermined.Glucoseandoleicacidmetabolismwerestudiedusingradio-labeled substrates,andgeneexpressionwasanalyzedusingreal-timeRT-PCR.Mitochondrialcontentandfunctionweremeasured by live imaging and determination of citrate synthase activity, respectively. Protein expression was assessed by electrophoresisandimmunoblotting. Results:High-frequency,acuteEPSincreaseddeoxyglucoseuptakeandlactateproduction,whilecellcontentsofbothATP andPCrdecreased.Chronic,low-frequencyEPSincreasedoxidativecapacityofculturedmyotubesbyincreasingglucose metabolism(uptakeandoxidation)andcompletefattyacidoxidation.mRNAexpressionlevelofpyruvatedehydrogenase complex4(PDK4)wassignificantlyincreasedinEPS-treatedcells,whilemRNAexpressionsofinterleukin6(IL-6),cytochrome Candcarnitinpalmitoyltransferaseb(CPT1b)alsotendedtoincrease.IntensityofMitoTrackerHRedFMwasdoubledafter 48hofchronic,low-frequencyEPS.Proteinexpressionofaslowfibertypemarker(MHCI)wasincreasedinEPS-treatedcells. Conclusions:OurresultsimplythatinvitroEPS(acute,high-frequentaswellaschronic,low-frequent)ofhumanmyotubes maybeusedtostudyeffectsofexercise. Citation:Nikolic′N,SkaretBakkeS,Tra