Intermedin inhibits vascular calcification by increasing the level of matrix gamma-carboxyglutamic acid protein.

AIMS Vascular calcification (VC) is highly associated with increased morbidity and mortality in patients with advanced chronic kidney disease. Paracrine/autocrine factors such as vasoactive peptides are involved in VC development. Here, we investigated the expression of the novel peptide intermedin (IMD) in the vasculature, tested its ability to prevent VC in vivo and in vitro, and examined the mechanism involved. METHODS AND RESULTS Rat VC was induced by administration of vitamin D3 plus nicotine (VDN). IMD (100 ng kg(-1) h(-1)) was systemically administered by a mini-osmotic pump. VDN-treated rat aortas showed lower IMD content and increased expression of its receptors, along with increased vascular calcium deposition and alkaline phosphatase (ALP) activity. Low IMD levels were accompanied by increased calcium deposition in human atherosclerotic plaques. In vivo administration of IMD greatly reduced vascular calcium deposition and ALP activity in VDN-treated rats when compared with vehicle treatment, which was further confirmed in cultured vascular smooth muscle cells. Concurrently, the loss of smooth-muscle lineage markers and matrix gamma-carboxyglutamic acid (Gla) protein (cMGP) in aortas was ameliorated by administering IMD to rats with VC, and the increased phosphor-Smad(1/5/8) and core binding factor alpha-1 levels in calcified vasculature were also reduced. However, the inhibitory effects of IMD on VC were eliminated upon pre-treatment with warfarin or small interfering RNA to reduce cMGP. CONCLUSION Reduced endogenous IMD levels are associated with increased mineralization in vivo, and administration of IMD inhibits VC development by increasing cMGP levels. IMD may be an endogenous vasoprotective factor for VC.

[1]  Yin Tintut,et al.  Vascular calcification: pathobiology of a multifaceted disease. , 2008, Circulation.

[2]  K. Boström,et al.  Proline and &ggr;-Carboxylated Glutamate Residues in Matrix Gla Protein Are Critical for Binding of Bone Morphogenetic Protein-4 , 2008, Circulation research.

[3]  B. McDermott,et al.  Intermedin (adrenomedullin‐2): a novel counter‐regulatory peptide in the cardiovascular and renal systems , 2008, British journal of pharmacology.

[4]  P. Kostenuik,et al.  Osteoprotegerin Inhibits Vascular Calcification Without Affecting Atherosclerosis in ldlr(−/−) Mice , 2008, Circulation.

[5]  L. Schurgers,et al.  Post‐translational modifications regulate matrix Gla protein function: importance for inhibition of vascular smooth muscle cell calcification , 2007, Journal of thrombosis and haemostasis : JTH.

[6]  Z. Al-Aly,et al.  Aortic Msx2-Wnt Calcification Cascade Is Regulated by TNF-&agr;–Dependent Signals in Diabetic Ldlr−/− Mice , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[7]  R. Franceschi,et al.  Regulation of matrix Gla protein by parathyroid hormone in MC3T3‐E1 osteoblast‐like cells involves protein kinase A and extracellular signal‐regulated kinase pathways , 2007, Journal of cellular biochemistry.

[8]  L. Schurgers,et al.  Undercarboxylated Matrix GLA Protein Levels Are Decreased in Dialysis Patients and Related to Parameters of Calcium-Phosphate Metabolism and Aortic Augmentation Index , 2007, Blood Purification.

[9]  J. Loscalzo,et al.  Vascular Calcification: Pathobiological Mechanisms and Clinical Implications , 2006, Circulation research.

[10]  K. Hruska,et al.  Function and effect of bone morphogenetic protein‐7 in kidney bone and the bone‐vascular links in chronic kidney disease , 2006, European journal of clinical investigation.

[11]  D. Towler,et al.  Molecular mechanisms of vascular calcification: lessons learned from the aorta. , 2006, Arteriosclerosis, thrombosis, and vascular biology.

[12]  Y. Ueta,et al.  Adrenomedullin 2 protects rat cerebral endothelial cells from oxidative damage in vitro , 2006, Brain Research.

[13]  Kazuo Kitamura,et al.  Adrenomedullin: A Protective Factor for Blood Vessels , 2005, Arteriosclerosis, thrombosis, and vascular biology.

[14]  G. London,et al.  Arteriosclerosis, vascular calcifications and cardiovascular disease in uremia , 2005, Current opinion in nephrology and hypertension.

[15]  B. Geng,et al.  Adrenomedullin(27–52) inhibits vascular calcification in rats , 2005, Regulatory Peptides.

[16]  C. Pan,et al.  Ghrelin blunted vascular calcification in vivo and in vitro in rats , 2005, Regulatory Peptides.

[17]  M. Mendelsohn,et al.  Angiotensin II and Aldosterone Regulate Gene Transcription Via Functional Mineralocortocoid Receptors in Human Coronary Artery Smooth Muscle Cells , 2005, Circulation research.

[18]  C. Pan,et al.  Effects of intermedin1–53 on cardiac function and ischemia/reperfusion injury in isolated rat hearts , 2005 .

[19]  Vincent Ouellette,et al.  Phenotypic Modulation of Vascular Smooth Muscle Cells During Medial Arterial Calcification: a Role for Endothelin? , 2004, Journal of cardiovascular pharmacology.

[20]  J. Mehta,et al.  Regulatory role of endothelium in the expression of genes affecting arterial calcification. , 2004, Biochemical and biophysical research communications.

[21]  C. Pan,et al.  The role of adrenomedullin and its receptor system in cardiovascular calcification of rat induced by Vitamin D3 plus nicotine , 2004, Peptides.

[22]  C. Klein,et al.  Intermedin Is a Calcitonin/Calcitonin Gene-related Peptide Family Peptide Acting through the Calcitonin Receptor-like Receptor/Receptor Activity-modifying Protein Receptor Complexes* , 2004, Journal of Biological Chemistry.

[23]  R. M. Pilkey,et al.  Cardioprotective medication use in hemodialysis patients. , 2003, The Canadian journal of cardiology.

[24]  Zhisheng Jiang,et al.  Effects of Adrenomedullin, C‐type Natriuretic Peptide, and Parathyroid Hormone‐Related Peptide on Calcification in Cultured Rat Vascular Smooth Muscle Cells , 2003, Journal of cardiovascular pharmacology.

[25]  P. Weissberg,et al.  Osteo/Chondrocytic Transcription Factors and Their Target Genes Exhibit Distinct Patterns of Expression in Human Arterial Calcification , 2003, Arteriosclerosis, thrombosis, and vascular biology.

[26]  N. Chen,et al.  Uremia induces the osteoblast differentiation factor Cbfa1 in human blood vessels. , 2003, Kidney international.

[27]  A. Sweatt,et al.  Matrix Gla protein (MGP) and bone morphogenetic protein‐2 in aortic calcified lesions of aging rats , 2003, Journal of thrombosis and haemostasis : JTH.

[28]  R. Aebersold,et al.  Smooth Muscle Cell Phenotypic Transition Associated With Calcification: Upregulation of Cbfa1 and Downregulation of Smooth Muscle Lineage Markers , 2001, Circulation research.

[29]  M. Engelse,et al.  Vascular calcification: expression patterns of the osteoblast-specific gene core binding factor alpha-1 and the protective factor matrix gla protein in human atherogenesis. , 2001, Cardiovascular research.

[30]  D. Sane,et al.  Arterial calcification: A review of mechanisms, animal models, and the prospects for therapy , 2001, Medicinal research reviews.

[31]  L. Demer,et al.  Matrix GLA Protein Modulates Differentiation Induced by Bone Morphogenetic Protein-2 in C3H10T1/2 Cells* , 2001, The Journal of Biological Chemistry.

[32]  J Rosing,et al.  Total chemical synthesis of human matrix Gla protein , 2001, Protein science : a publication of the Protein Society.

[33]  D. Sane,et al.  Modulation of the Binding of Matrix Gla Protein (MGP) to Bone Morphogenetic Protein-2 (BMP-2) , 2000, Thrombosis and Haemostasis.

[34]  P. Price,et al.  Warfarin-induced artery calcification is accelerated by growth and vitamin D. , 2000, Arteriosclerosis, thrombosis, and vascular biology.

[35]  T. Nakata,et al.  Hypoparathyroidism Potentiates Cardiovascular Complications through Disturbed Calcium Metabolism: Possible Risk of Vitamin D3 Analog Administration in Dialysis Patients with End-Stage Renal Disease , 2000, Nephron.

[36]  I. Shapiro,et al.  Matrix Gla Protein Is a Developmental Regulator of Chondrocyte Mineralization And, When Constitutively Expressed, Blocks Endochondral and Intramembranous Ossification in the Limb , 1999, The Journal of cell biology.

[37]  Y. Nishizawa,et al.  1,25-Dihydroxyvitamin D3 increases in vitro vascular calcification by modulating secretion of endogenous parathyroid hormone-related peptide. , 1998, Circulation.

[38]  YoshikiNishizawa,et al.  1,25-Dihydroxyvitamin D3 Increases In Vitro Vascular Calcification by Modulating Secretion of Endogenous Parathyroid Hormone–Related Peptide , 1998 .

[39]  P. Price,et al.  Warfarin causes rapid calcification of the elastic lamellae in rat arteries and heart valves. , 1998, Arteriosclerosis, thrombosis, and vascular biology.

[40]  P. Weissberg,et al.  Vascular calcification: new insights into an old problem , 1998, The Journal of pathology.

[41]  J. Skepper,et al.  Calcification of human vascular cells in vitro is correlated with high levels of matrix Gla protein and low levels of osteopontin expression. , 1998, Arteriosclerosis, thrombosis, and vascular biology.

[42]  J. Atkinson,et al.  Aortic calcification produced by vitamin D3 plus nicotine. , 1997, Journal of vascular research.

[43]  R. Behringer,et al.  Spontaneous calcification of arteries and cartilage in mice lacking matrix GLA protein , 1997, Nature.

[44]  H. Koyama,et al.  β-Glycerophosphate Accelerates Calcification in Cultured Bovine Vascular Smooth Muscle Cells , 1995 .

[45]  P. Price,et al.  Primary structure of bovine matrix Gla protein, a new vitamin K-dependent bone protein. , 1985, The Journal of biological chemistry.

[46]  M. Urist,et al.  Matrix Gla protein, a new gamma-carboxyglutamic acid-containing protein which is associated with the organic matrix of bone. , 1983, Biochemical and biophysical research communications.

[47]  A. Parfitt.  Soft-tissue calcification in uremia. , 1969, Archives of internal medicine.

[48]  K. Hruska,et al.  Reversal of the adynamic bone disorder and decreased vascular calcification in chronic kidney disease by sevelamer carbonate therapy. , 2007, Journal of the American Society of Nephrology : JASN.

[49]  C. Pan,et al.  Effects of intermedin(1-53) on cardiac function and ischemia/reperfusion injury in isolated rat hearts. , 2005, Biochemical and biophysical research communications.

[50]  P. Weissberg,et al.  The role of Gla proteins in vascular calcification. , 1998, Critical reviews in eukaryotic gene expression.