Menaquinone-4 modulates the expression levels of calcification-associated factors to inhibit calcification of rat aortic vascular smooth muscle cells in a dose-dependent manner.

Vascular calcification (VC) caused by chronic kidney disease (CKD)-mineral and bone disorder is a common complication of CKD. Recent studies have demonstrated that menaquinone-4 (MK-4) is negativly associated with VC in patients with CKD. Furthermore, we have previously shown that runt-related transcription factor 2 (Runx2) is important in the phenotypic transformation process of rat vascular smooth muscle cells (VSMCs), which is the key step for the development of VC. The present study investigated the influence of MK-4 on the phenotypic transformation process of rat VSMCs in order to illustrate its role in the process of VC. Calcification assays were perfomed to access the calcified degree of rat VSMCs. Additionally, the genes and proteins related to phenotypic transformation were measured by reverse transcription-polymerase chain reaction and western blotting methods. It was revealed that calcium deposition in the cells was evidently increased with an addition of β-glycerophosphate (β-GP) and could be completely prevented by co-incubation with MK-4 in a dose-dependent manner. Furthermore, the expression of Runx2 in the β-GP-induced VSMCs was inhibited by MK-4. It was also revealed that the expression of SMAD1 and bone morphogenetic protein (BMP)-2 were decreased in the β-GP-induced VSMCs treated with MK-4 in a dose-dependent manner; however, the expression of SMAD7 was increased in the β-GP-induced VSMCs treated with MK-4 in a dose-dependent manner. These observations suggest that MK-4 reduces mineralization by regulating the BMP-2 signaling pathway in order to attenuate the expression of Runx2.

[1]  Junxia Zhang,et al.  Extracellular acidosis suppresses calcification of vascular smooth muscle cells by inhibiting calcium influx via L-type calcium channels , 2018, Clinical and experimental hypertension.

[2]  C. Heizmann,et al.  Extracellular S100A4 induces smooth muscle cell phenotypic transition mediated by RAGE. , 2015, Biochimica et biophysica acta.

[3]  C. Giachelli,et al.  Runx2 Expression in Smooth Muscle Cells Is Required for Arterial Medial Calcification in Mice. , 2015, The American journal of pathology.

[4]  Jiliang Wu,et al.  Losartan Inhibits Vascular Calcification by Suppressing the BMP2 and Runx2 Expression in Rats In Vivo , 2015, Cardiovascular Toxicology.

[5]  Junxia Zhang,et al.  Alteration of Type I Collagen in the Radial Artery of Patients With End-Stage Renal Disease , 2015, The American journal of the medical sciences.

[6]  Junxia Zhang,et al.  Magnesium modulates the expression levels of calcification-associated factors to inhibit calcification in a time-dependent manner , 2015, Experimental and therapeutic medicine.

[7]  A. H. Campos,et al.  BMP-2 and -4 produced by vascular smooth muscle cells from atherosclerotic lesions induce monocyte chemotaxis through direct BMPRII activation. , 2014, Atherosclerosis.

[8]  B. Yu,et al.  BMP-2 and titanium particles synergistically activate osteoclast formation , 2014, Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas.

[9]  M. Wan,et al.  Mesenchymal stem cells recruited by active TGFβ contribute to osteogenic vascular calcification. , 2014, Stem cells and development.

[10]  D. Hemmings,et al.  Review: novel insights into the regulation of vascular tone by sphingosine 1-phosphate. , 2014, Placenta.

[11]  Noriko KoitayaMariko,et al.  Low-dose vitamin K 2 (MK-4) supplementation for 12 months improves bone metabolism and prevents forearm bone loss in postmenopausal Japanese women , 2014 .

[12]  M. Wahlqvist,et al.  Clinical decision-making for vitamin K-1 and K-2 deficiency and coronary artery calcification with warfarin therapy: are diet, factor Xa inhibitors or both the answer? , 2013, Asia Pacific journal of clinical nutrition.

[13]  H. Olauson,et al.  FGF23 and Klotho in chronic kidney disease , 2013, Current opinion in nephrology and hypertension.

[14]  J. Uitto,et al.  Vitamin K-dependent carboxylation of matrix Gla-protein: a crucial switch to control ectopic mineralization. , 2013, Trends in molecular medicine.

[15]  J. Ix,et al.  Clinical detection, risk factors, and cardiovascular consequences of medial arterial calcification: a pattern of vascular injury associated with aberrant mineral metabolism. , 2013, Seminars in nephrology.

[16]  N. Cherbuin.  Higher dietary intakes of potassium, calcium and magnesium are associated with a reduced risk of developing vascular dementia , 2012, Evidence-Based Mental Health.

[17]  P. ten Dijke,et al.  BMP signaling in vascular diseases , 2012, FEBS letters.

[18]  L. Schurgers,et al.  Effect of vitamin K2 supplementation on functional vitamin K deficiency in hemodialysis patients: a randomized trial. , 2012, American journal of kidney diseases : the official journal of the National Kidney Foundation.

[19]  P. Bolasco.  Effects of the Use of Non-Calcium Phosphate Binders in the Control and Outcome of Vascular Calcifications: A Review of Clinical Trials on CKD Patients , 2011, International journal of nephrology.

[20]  L. Schurgers,et al.  Circulating matrix γ‐carboxyglutamate protein (MGP) species are refractory to vitamin K treatment in a new case of Keutel syndrome , 2011, Journal of thrombosis and haemostasis : JTH.

[21]  L. Schurgers,et al.  Biosynthesis of the vitamin K-dependent matrix Gla protein (MGP) in chondrocytes: a fetuin-MGP protein complex is assembled in vesicles shed from normal but not from osteoarthritic chondrocytes. , 2010, Osteoarthritis and cartilage.

[22]  S. Booth,et al.  Vitamins K and D status in stages 3-5 chronic kidney disease. , 2010, Clinical journal of the American Society of Nephrology : CJASN.

[23]  Brian J. Bennett,et al.  Inhibition of bone morphogenetic protein protects against atherosclerosis and vascular calcification , 2010, Circulation research.

[24]  Kohei Miyazono,et al.  Bone morphogenetic protein receptors and signal transduction. , 2010, Journal of biochemistry.

[25]  C. Giachelli.  The emerging role of phosphate in vascular calcification. , 2009, Kidney international.

[26]  T. Adragão.  Evaluation of Vascular Calcifications in CKD Patients , 2009, The International journal of artificial organs.

[27]  S. Sasaki,et al.  Effect of low dose vitamin K2 (MK-4) supplementation on bio-indices in postmenopausal Japanese women. , 2009, Journal of nutritional science and vitaminology.

[28]  P. Kerr,et al.  Mortality risk for dialysis patients with different levels of serum calcium, phosphorus, and PTH: the Dialysis Outcomes and Practice Patterns Study (DOPPS). , 2008, American journal of kidney diseases : the official journal of the National Kidney Foundation.

[29]  S. Booth,et al.  Vitamin K Status of Canadian Peritoneal Dialysis Patients , 2008, Peritoneal dialysis international : journal of the International Society for Peritoneal Dialysis.

[30]  Su‐Li Cheng,et al.  Vascular Bmp–Msx2–Wnt Signaling and Oxidative Stress in Arterial Calcification , 2007, Annals of the New York Academy of Sciences.

[31]  R. Pereira,et al.  Bone metabolism and vascular calcification. , 2007, Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas.

[32]  A. Garg,et al.  Chronic kidney disease and mortality risk: a systematic review. , 2006, Journal of the American Society of Nephrology : JASN.

[33]  Xianwu Li,et al.  Role of the Sodium-Dependent Phosphate Cotransporter, Pit-1, in Vascular Smooth Muscle Cell Calcification , 2006, Circulation research.

[34]  A. Hofman,et al.  Dietary intake of menaquinone is associated with a reduced risk of coronary heart disease: the Rotterdam Study. , 2004, The Journal of nutrition.

[35]  H. Sakai,et al.  Upregulation of rhoA mRNA in bronchial smooth muscle of antigen-induced airway hyperresponsive rats. , 2003, Journal of smooth muscle research = Nihon Heikatsukin Gakkai kikanshi.

[36]  L. Schurgers,et al.  Tissue-Specific Utilization of Menaquinone-4 Results in the Prevention of Arterial Calcification in Warfarin-Treated Rats , 2003, Journal of Vascular Research.

[37]  Jay R Lieberman,et al.  The role of growth factors in the repair of bone. Biology and clinical applications. , 2002, The Journal of bone and joint surgery. American volume.

[38]  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.

[39]  E. Brown,et al.  Extracellular calcium elicits a chemokinetic response from monocytes in vitro and in vivo. , 2000, The Journal of clinical investigation.

[40]  K. Miyazono,et al.  Signal transduction by bone morphogenetic proteins. , 1998, Cytokine & growth factor reviews.

[41]  J. Campbell,et al.  Culture techniques and their applications to studies of vascular smooth muscle. , 1993, Clinical science.