Calcification inhibitors and Wnt signaling proteins are implicated in bovine artery smooth muscle cell calcification in the presence of phosphate and vitamin D sterols.
Administration of active vitamin D sterols to treat secondary hyperparathyroidism in patients with chronic kidney disease receiving dialysis has been associated with elevated serum calcium and phosphorus levels, which may lead to increased risk of vascular calcification. However, calcimimetics, by binding to the parathyroid gland calcium-sensing receptors, reduce serum parathyroid hormone, calcium, phosphorus, and the calcium-phosphorus product. Using cultured bovine aorta vascular smooth muscle cells (BASMCs), an in vitro model of vascular calcification, we compared calcification levels and gene expression profiles after exposure to the phosphate source ss-glycerolphosphate (BGP), the active vitamin D sterols calcitriol and paricalcitol, the calcimimetic R-568, or BGP with the active vitamin D sterols or R-568. Cells exposed to BGP (10 mM) alone or with calcitriol or paricalcitol showed dose-dependent BASMC calcification. No change in calcification was observed in cultures exposed to BGP with R-568, consistent with the observed lack of calcium-sensing receptor expression. Microarray analysis using total cellular RNA from cultures exposed to vehicle or BGP in the absence and presence of 10(-8) M calcitriol or paricalcitol for 7 days showed that cells exposed to BGP with calcitriol or BGP with paricalcitol had virtually identical gene expression profiles, which differed from those of cells treated with BGP or vehicle alone. Several osteoblast- and chondrocyte-associated genes were modulated by BGP and vitamin D exposure. In this study, exposure of BASMCs to phosphate and active vitamin D sterols induced calcification and changes in expression of genes associated with mineralized tissue.
Shalhoub
V
V
Department of Metabolic Disorders, Amgen, Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USA. shalhoub@amgen.com
Shatzen
E
E
Henley
C
C
Boedigheimer
M
M
McNinch
J
J
Manoukian
R
R
Damore
M
M
Fitzpatrick
D
D
Haas
K
K
Twomey
B
B
Kiaei
P
P
Ward
S
S
Lacey
D L
DL
Martin
D
D
D000469:Alkaline Phosphatase / Q000378:metabolism; D000814:Aniline Compounds / Q000494:pharmacology*; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002114:Calcinosis / Q000139:chemically induced / Q000378:metabolism / Q000517:prevention & control*; D002117:Calcitriol / Q000494:pharmacology*; D002118:Calcium / Q000819:agonists / Q000378:metabolism / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D004338:Drug Combinations; D004872:Ergocalciferols / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D005994:Glycerophosphates / Q000494:pharmacology*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D010627:Phenethylamines; D010758:Phosphorus / Q000378:metabolism / Q000494:pharmacology; D011437:Propylamines; D018167:Receptors, Calcitriol / Q000235:genetics / Q000378:metabolism; D044169:Receptors, Calcium-Sensing / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D051153:Wnt Proteins / Q000502:physiology*
D000814:Aniline Compounds; D004338:Drug Combinations; D004872:Ergocalciferols; D005994:Glycerophosphates; C107873:N-(2-chlorophenylpropyl)-1-(3-methoxyphenyl)ethylamine; D010627:Phenethylamines; D011437:Propylamines; D018167:Receptors, Calcitriol; D044169:Receptors, Calcium-Sensing; D051153:Wnt Proteins; D010758:Phosphorus; C084656:paricalcitol; D000469:Alkaline Phosphatase; D002117:Calcitriol; D002118:Calcium