caa a22 4500 606235795 CHVBK 20210128101244.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s11010-015-2552-6 doi (NATIONALLICENCE)springer-10.1007/s11010-015-2552-6 A new in vitro model to delay high phosphate-induced vascular calcification progression [Elektronische Daten] [Paola Ciceri, Francesca Elli, Laura Cappelletti, Delfina Tosi, Paola Braidotti, Gaetano Bulfamante, Mario Cozzolino] An increasing amount of patients affected by advanced chronic kidney disease suffer from vascular calcification (VC) that associates with cardiovascular morbidity and mortality. In this study, we created a new experimental in vitro model, trying to better elucidate high phosphate (Pi)-induced VC pathogenic mechanisms. Rat aortic vascular smooth muscle cells (VSMCs) were challenged for 7-10days with high Pi with a repeated and short suspensions of high Pi treatment (intermittent suspension, IS) that was able to induce a significant inhibition of high Pi calcification, maximal at 5h. Interestingly, the delay in calcification is a consequence of either the absence of free Pi or calcium-phosphate crystals being comparable to the total effect obtained during the 5h-IS. The protective effect of IS was mediated by the reduction of apoptosis as demonstrated by the action of 20μmol/L Z-VAD-FMK and by the preservation of the pro-survival receptor Axl expression. Furthermore, autophagy, during IS, was potentiated by increasing the autophagic flux, evaluated by LC3IIB western, while treating VSMCs with 1mmol/L valproic acid did not affect VC. Finally, IS prevented VSMC osteoblastic differentiation by preserving smooth muscle lineage markers expression. Our data support the hypothesis that to delay significantly VC is necessary and sufficient the IS of high Pi challenge. The IS was able to prevent significantly apoptosis, to induce a potentiation in autophagy, and to prevent osteoblastic differentiation by preserving SM lineage markers. Springer Science+Business Media New York, 2015 Vascular calcification nationallicence Apoptosis nationallicence VSMC nationallicence Phosphate nationallicence Ciceri Paola Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut Elli Francesca Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut Cappelletti Laura Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut Tosi Delfina Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut Braidotti Paola Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut Bulfamante Gaetano Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut Cozzolino Mario Renal Division and Laboratory of Experimental Nephrology, Dipartimento di Scienze della Salute, Università di Milano, S. Paolo Hospital, Milan, Via A. di Rudinì, 8, 20142, Milan, Italy aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 410/1-2(2015-12-01), 197-206 0300-8177 410:1-2<197 2015 410 11010 https://doi.org/10.1007/s11010-015-2552-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-015-2552-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ciceri Paola Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut NATIONALLICENCE 700 1- Elli Francesca Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut NATIONALLICENCE 700 1- Cappelletti Laura Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut NATIONALLICENCE 700 1- Tosi Delfina Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut NATIONALLICENCE 700 1- Braidotti Paola Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut NATIONALLICENCE 700 1- Bulfamante Gaetano Laboratory of Experimental Nephrology, Department of Health Sciences, University of Milan, Milan, Italy aut NATIONALLICENCE 700 1- Cozzolino Mario Renal Division and Laboratory of Experimental Nephrology, Dipartimento di Scienze della Salute, Università di Milano, S. Paolo Hospital, Milan, Via A. di Rudinì, 8, 20142, Milan, Italy aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 410/1-2(2015-12-01), 197-206 0300-8177 410:1-2<197 2015 410 11010