Calcimimetic R-568 and Its Enantiomer S-568 Increase Nitric Oxide Release in Human Endothelial Cells 英文参考文献.docVIP

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Calcimimetic R-568 and Its Enantiomer S-568 Increase Nitric Oxide Release in Human Endothelial Cells 英文参考文献.doc

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Calcimimetic R-568 and Its Enantiomer S-568 Increase Nitric Oxide Release in Human Endothelial Cells 英文参考文献

CalcimimeticR-568andItsEnantiomerS-568Increase NitricOxideReleaseinHumanEndothelialCells MarioBonomini1,4.,AnnalisaGiardinelli2,5.,CaterinaMorabito3,5,SaraDiSilvestre2,5,MorenoDi Cesare1,4,NataliaDiPietro2,5,VittorioSirolli1,4,GloriaFormoso4,5,LuigiAmoroso1,4,MariaAddolorata Mariggio`3,5,AssuntaPandolfi2,5 * 1Institute of Nephrology-Department of Medicine, University ‘‘G. d’Annunzio’’, Chieti-Pescara, Italy, 2Department of Biomedical Sciences, University ‘‘G. d’Annunzio’’, Chieti-Pescara, Italy, 3Department of Neuroscience and Imaging, University ‘‘G. d’Annunzio’’, Chieti-Pescara, Italy, 4Department of Medicine and Aging Sciences, University ‘‘G. d’Annunzio’’, Chieti-Pescara, Italy, 5Aging Research Center, Centro Scienze dell’Invecchiamento (Ce.S.I.), ‘‘G. d’Annunzio’’ University Foundation, Chieti- Pescara,Italy Abstract Background: Calcimimetics, such as R-568, are thought to activate G protein-linked Ca2+-sensing receptor (CaSR) by allosterically increasing the affinity of the receptor for Ca2+ allowing for efficient control of uremic hyperparathyroidism. Severalrecentstudiessuggesttheypossessadditionalvascularactions.Althoughithasbeenpostulatedthatcalcimimetics mayhaveadirecteffectonCaSRinthebloodvessels,furtherstudiesareneededtoelucidatetheirvascularCaSR-dependent versusCaSR-independenteffects. Methodology/Principal Findings: Focusing on human umbilical vein endothelial cells (HUVECs), we studied the CaSR expressionanddistributionbyImmunofluorescenceandWesternBlotanalysis.CaSRfunctionwasevaluatedbymeasuring thepotentialeffectofcalcimimeticR-568anditsenantiomerS-568uponthemodulationofintracellularCa2+levels(usinga singlecellapproachandFURA-2AM),inthepresenceorabsenceofCalhex-231,anegativemodulatorofCaSR.Toaddress theirpotentialvascularfunctions,wealsoevaluatedR-andS-568-stimulatedenzymaticreleaseofNitricOxide(NO)byDAF- 2DA, by Nitric Oxide Synthase (NOS) radiometric assay (both in HUVECs and in Human Aortic Endothelial Cells) and by measuring eNOS-ser1177 phos