Moderate Exercise Promotes Human RBC-NOS Activity, NO Production and Deformability through Akt Kinase Pathway 英文参考文献.docVIP

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Moderate Exercise Promotes Human RBC-NOS Activity, NO Production and Deformability through Akt Kinase Pathway 英文参考文献.doc

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Moderate Exercise Promotes Human RBC-NOS Activity, NO Production and Deformability through Akt Kinase Pathway 英文参考文献

ModerateExercisePromotesHumanRBC-NOSActivity, NOProductionandDeformabilitythroughAktKinase Pathway FrankSuhr1,2,JulianBrenig1,RebeccaMu¨ller1,HilkeBehrens1,WilhelmBloch1,2,MarijkeGrau1* 1DepartmentofMolecularandCellularSportsMedicine,GermanSportUniversityCologne,Cologne,Germany, 2TheGermanResearchCenterofEliteSport,German SportUniversityCologne,Cologne,Germany Abstract Background: Nitric oxide (NO) produced by nitric oxide synthase (NOS) in human red blood cells (RBCs) was shown to dependonshearstressandtoexhibitimportantbiologicalfunctions,suchasinhibitionofplateletactivation.Inthepresent study we hypothesized that exercise-induced shear stress stimulates RBC-NOS activation pathways, NO signaling, and deformabilityofhumanRBCs. Methods/Findings:Fifteenmalesubjectsconductedanexercisetestwithvenousbloodsamplingbeforeandafterrunning on a treadmill for 1 hour. Immunohistochemical staining as well as western blot analysis were used to determine phosphorylationandthusactivationofAktkinaseandRBC-NOSaswellasaccumulationofcyclicguanylylmonophosphate (cGMP) induced by the intervention. The data revealed that activation of NO upstream located enzyme Akt kinase was significantly increased after the test. Phosphorylation of RBC-NOSSer1177 was also significantly increased after exercise, indicating activation of RBC-NOS through Akt kinase. Total detectable RBC-NOS content and phosphorylation of RBC- NOSThr495 were not affected by the intervention. NO production by RBCs, determined by DAF fluorometry, and RBC deformability, measured via laser-assisted-optical-rotational red cell analyzer, were also significantly increased after the exercise test. The content of the NO downstream signaling molecule cGMP increased after the test. Pharmacological inhibitionofphosphatidylinositol3(PI3)-kinase/AktkinasepathwayledtoadecreaseinRBC-NOSactivation,NOproduction andRBCdeformability. Conclusion/Significance: Thishumaninvivostudyfirst-timeprovidesstrongevidencethatexercise-inducedshear