The mechanisms of uremic serum-induced expression of bone matrix proteins in bovine vascular smooth muscle cells

Xuening (Neal) Chen, D. Duan, K. D. O'Neill, G. O. Wolisi, J. J. Koczman, R. LaClair, Sharon Moe

Research output: Contribution to journalArticle

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Abstract

We have previously found that uremic human serum upregulates RUNX2 in vascular smooth muscle cells (VSMCs), and that RUNX2 is upregulated in areas of vascular calcification in vivo. To confirm the role of RUNX2, we transiently transfected a dominant-negative RUNX2 (ΔRUNX2) construct in bovine vascular smooth muscle cells (BVSMCs). Blocking RUNX2 transcriptional activity significantly decreased uremic serum induced alkaline phosphatase (ALP) activity (268 ± 34 vs 188 ± 9.5 U/g protein, P < 0.05) and osteocalcin expression (172 ± 17 vs 125 ± 9 ODU, P < 0.05). To determine the mechanism by which uremic serum upregulates RUNX2, we examined cell signaling pathways. BVSMCs were incubated in the presence or absence of inhibitors and RUNX2 expression and ALP activity were determined. The results demonstrate that the cyclic AMP (cAMP)/protein kinase A (PKA), but not protein kinase C, signaling pathway is involved in uremic serum-induced RUNX2 expression and ALP activity in BVSMCs. To examine potential uremic 'toxins', we measured bone morphogenetic protein (BMP)-2 concentration and found that uremic serum contained increased BMP-2 (uremic serum = 169 ± 33 pg/ml, normal serum = 117 ± 15 pg/ml, P < 0.05). The incubation of BVSMCs with noggin, an inhibitor of BMP, decreased RUNX2 expression. In addition, BMP-2 secretion progressively increased during calcification and uremic serum enhanced its secretion compared to normal serum. In conclusion, this study demonstrates that RUNX2 transcriptional activity is critical in uremic serum-induced bone matrix protein expression in BVSMCs and that the cAMP/PKA pathway is involved. BMP-2 is also increased in uremic serum and can upregulate RUNX2 and calcification in vitro in VSMCs.

Original languageEnglish
Pages (from-to)1046-1053
Number of pages8
JournalKidney International
Volume70
Issue number6
DOIs
StatePublished - Sep 2006

Fingerprint

Bone Matrix
Vascular Smooth Muscle
Smooth Muscle Myocytes
Serum
Bone Morphogenetic Protein 2
Proteins
Alkaline Phosphatase
Adenylate Kinase
Up-Regulation
Cyclic AMP-Dependent Protein Kinases
Cyclic AMP
Vascular Calcification
Bone Morphogenetic Proteins
Osteocalcin
Protein Kinase C

Keywords

  • Bone morphogenetic protein 2 (BMP-2)
  • Chronic kidney disease (CKD)
  • Core binding factor α1 (RUNX2)
  • Vascular calcification

ASJC Scopus subject areas

  • Nephrology

Cite this

The mechanisms of uremic serum-induced expression of bone matrix proteins in bovine vascular smooth muscle cells. / Chen, Xuening (Neal); Duan, D.; O'Neill, K. D.; Wolisi, G. O.; Koczman, J. J.; LaClair, R.; Moe, Sharon.

In: Kidney International, Vol. 70, No. 6, 09.2006, p. 1046-1053.

Research output: Contribution to journalArticle

Chen, Xuening (Neal) ; Duan, D. ; O'Neill, K. D. ; Wolisi, G. O. ; Koczman, J. J. ; LaClair, R. ; Moe, Sharon. / The mechanisms of uremic serum-induced expression of bone matrix proteins in bovine vascular smooth muscle cells. In: Kidney International. 2006 ; Vol. 70, No. 6. pp. 1046-1053.
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AU - Duan, D.

AU - O'Neill, K. D.

AU - Wolisi, G. O.

AU - Koczman, J. J.

AU - LaClair, R.

AU - Moe, Sharon

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AB - We have previously found that uremic human serum upregulates RUNX2 in vascular smooth muscle cells (VSMCs), and that RUNX2 is upregulated in areas of vascular calcification in vivo. To confirm the role of RUNX2, we transiently transfected a dominant-negative RUNX2 (ΔRUNX2) construct in bovine vascular smooth muscle cells (BVSMCs). Blocking RUNX2 transcriptional activity significantly decreased uremic serum induced alkaline phosphatase (ALP) activity (268 ± 34 vs 188 ± 9.5 U/g protein, P < 0.05) and osteocalcin expression (172 ± 17 vs 125 ± 9 ODU, P < 0.05). To determine the mechanism by which uremic serum upregulates RUNX2, we examined cell signaling pathways. BVSMCs were incubated in the presence or absence of inhibitors and RUNX2 expression and ALP activity were determined. The results demonstrate that the cyclic AMP (cAMP)/protein kinase A (PKA), but not protein kinase C, signaling pathway is involved in uremic serum-induced RUNX2 expression and ALP activity in BVSMCs. To examine potential uremic 'toxins', we measured bone morphogenetic protein (BMP)-2 concentration and found that uremic serum contained increased BMP-2 (uremic serum = 169 ± 33 pg/ml, normal serum = 117 ± 15 pg/ml, P < 0.05). The incubation of BVSMCs with noggin, an inhibitor of BMP, decreased RUNX2 expression. In addition, BMP-2 secretion progressively increased during calcification and uremic serum enhanced its secretion compared to normal serum. In conclusion, this study demonstrates that RUNX2 transcriptional activity is critical in uremic serum-induced bone matrix protein expression in BVSMCs and that the cAMP/PKA pathway is involved. BMP-2 is also increased in uremic serum and can upregulate RUNX2 and calcification in vitro in VSMCs.

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KW - Vascular calcification

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