title: Vascular calcification and inorganic phosphate.
abstract: Vascular calcification is highly correlated with elevated serum phosphate levels in uremic patients. To shed light on this process, we examined the ability of extracellular inorganic phosphate (Pi) levels to regulate human aortic smooth muscle cell (HSMC) culture mineralization in vitro. When cultured in media containing normal physiological levels of Pi (1.4 mmol/L Pi), HSMC grew in monolayers and did not mineralize. In contrast, HSMC cultured in media containing Pi levels comparable to those seen in hyperphosphatemic individuals (>1.4 mmol/L), showed dose-dependent increases in cell culture calcium deposition. Mechanistic studies showed that elevated Pi treatment of HSMC also enhanced the expression of the osteogenic markers, osteocalcin and Cbfa-1. The effects of elevated Pi on HSMC were mediated by a sodium-dependent phosphate cotransporter (NPC), as indicated by the ability of the specific NPC inhibitor, phosphonoformic acid (PFA), to dose-dependently inhibit Pi-induced calcium deposition as well as osteocalcin and Cbfa-1 gene expression. Using polymerase chain reaction and Northern blot analyses, the NPC in HSMC was identified as Pit-1 (Glvr-1), a member of the type III NPCs. Interestingly, platelet-derived growth factor-BB (PDGF-BB), a potent atherogenic stimulus, increased the maximum velocity (Vmax) but not the affinity (Km) of phosphate uptake, enhanced the expression of Pit-1 mRNA, and induced HSMC culture calcification in a time- and dose-dependent manner. Importantly, in the presence of PDGF, HSMC culture calcification occurred under normophosphatemic conditions. These data suggest that elevated Pi may directly stimulate HSMC to undergo phenotypic changes that predispose to calcification and may help explain both the phenomena of human metastatic calcification under hyperphosphatemic conditions as well as increased calcification in PDGF-rich atherosclerotic lesions.
authors.affiliation: Department of Bioengineering, University of Washington, Seattle, WA, USA. ceci@u.washington.edu
authors.forename: C M
authors.initials: CM
authors.lastname: Giachelli
authors.forename: S
authors.initials: S
authors.lastname: Jono
authors.forename: A
authors.initials: A
authors.lastname: Shioi
authors.forename: Y
authors.initials: Y
authors.lastname: Nishizawa
authors.forename: K
authors.initials: K
authors.lastname: Mori
authors.forename: H
authors.initials: H
authors.lastname: Morii
mesh_terms: D000199:Actins / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D002114:Calcinosis / Q000209:etiology* / Q000378:metabolism*; D002318:Cardiovascular Diseases / Q000209:etiology* / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D017245:Foscarnet / Q000494:pharmacology; D015870:Gene Expression; D005994:Glycerophosphates / Q000494:pharmacology; D006801:Humans; D008565:Membrane Proteins; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D009363:Neoplasm Proteins; D015675:Osteocalcin / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism / Q000493:pharmacokinetics*; D049294:Phospholipid Transfer Proteins; D010982:Platelet-Derived Growth Factor / Q000378:metabolism / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D050608:Sodium-Phosphate Cotransporter Proteins; D050611:Sodium-Phosphate Cotransporter Proteins, Type III; D027981:Symporters / Q000037:antagonists & inhibitors / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D014511:Uremia / Q000150:complications*
chemical_list: D000199:Actins; D002352:Carrier Proteins; D005994:Glycerophosphates; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D010710:Phosphates; D049294:Phospholipid Transfer Proteins; D010982:Platelet-Derived Growth Factor; D012333:RNA, Messenger; C493586:SLC20A1 protein, human; D050608:Sodium-Phosphate Cotransporter Proteins; D050611:Sodium-Phosphate Cotransporter Proteins, Type III; D027981:Symporters; D014157:Transcription Factors; D015675:Osteocalcin; D017245:Foscarnet