Role of PKC and CaV1.2 in Detrusor Overactivity in a Model of Obesity Associated with Insulin Resistance in Mice 英文参考文献.docVIP

RoleofPKCandCaV1.2inDetrusorOveractivityina ModelofObesityAssociatedwithInsulinResistancein Mice LuizO.Leiria1,CarolinaSollon1,MarinaC.Calixto1,Let?′ciaLintomen1,Fab?′olaZ.Mo′nica1, GabrielF.Anhe?1,GilbertoDeNucci1,AngelinaZanesco2,AndrewD.Grant3,EdsonAntunes1* 1DepartmentofPharmacology,FacultyofMedicalSciences,UniversityofCampinas(UNICAMP),Campinas,Sa?oPaulo,Brazil,2DepartmentofPhysicalEducation,Institute ofBioscience,UniversityofSa?o PauloState(UNESP),RioClaro,Sa?o Paulo,Brazil,3WolfsonCentreforAge-RelatedDiseases,King’sCollege,London,UnitedKingdom Abstract Obesity/metabolicsyndromearecommonriskfactorsforoveractivebladder.Thisstudyaimedtoinvestigatethefunctional andmolecularchangesofdetrusorsmoothmuscle(DSM)inhigh-fatinsulinresistantobesemice,focusingontheroleof proteinkinaseC (PKC)andCav1.2 incausing bladder dysfunction.MaleC57BL/6 mice werefedwithhigh-fatdietfor 10 weeks.Invitrofunctionalresponsesandcystometry,aswellasPKCandCa 1.2expressioninbladderwereevaluated.Obese v miceexhibitedhigherbodyweight,epididymalfatmass,fastingglucoseandinsulinresistance.Carbachol(0.001–100 mM), a,b-methylene ATP (1–10mM), KCl (1–300mM), extracellular Ca2+ (0.01–100mM) and phorbol-12,13-dibutyrate (PDBu; 0.001–3mM) all produced greater DSM contractions in obese mice, which were fully reversed by the Cav1.2 blocker amlodipine.Cystometryevidencedaugmentedfrequency,non-voidcontractionsandpost-voidpressureinobesemicethat were also prevented by amlodipine. Metformin treatment improved the insulin sensitivity, and normalized the in vitro bladderhypercontractilityandcystometricdysfunctioninobesemice.ThePKCinhibitorGF109203X(1 mM)alsoreducedthe carbacholinducedcontractions.PKCproteinexpressionwasmarkedlyhigherinbladdertissuesfromobesemice,whichwas normalizedbymetformintreatment.TheCav1.2channelproteinexpressionwasnotmodifiedinanyexperimentalgroup. Our findings show that Cav1.2 blockade and improvement of insulin sensitization restores the enhanced PKC protein exp