Augmentation of phosphate-induced osteo-/chondrogenic transformation of vascular smooth muscle cells by homoarginine.

AIMS Reduced homoarginine plasma levels are associated with unfavourable cardiovascular outcome in chronic kidney disease (CKD). Cardiovascular events in CKD are fostered by vascular calcification, an active process promoted by hyperphosphatemia and involving osteo-/chondrogenic transformation of vascular smooth muscle cells (VSMCs). The present study explored the effect of homoarginine on phosphate-induced osteo-/chondrogenic signalling and vascular calcification. METHODS AND RESULTS Experiments were performed in hyperphosphatemic klotho-hypomorphic mice (kl/kl), in subtotal nephrectomy and vitamin D3-overload mouse calcification models and in primary human aortic smooth muscle cells (HAoSMCs). As a result, plasma homoarginine levels were lower in kl/kl mice than in wild-type mice and in both genotypes significantly increased by lifelong treatment with homoarginine. Surprisingly, homoarginine treatment of kl/kl mice and of mice with renal failure after subtotal nephrectomy augmented vascular calcification and enhanced the transcript levels of plasminogen activator inhibitor 1 (Pai1) and of osteogenic markers Msx2, Cbfa1, and Alpl. Similarly, homoarginine treatment of HAoSMCs increased phosphate-induced calcium deposition, ALP activity, as well as PAI1, MSX2, CBFA1, and ALPL mRNA levels. Homoarginine alone up-regulated osteo-/chondrogenic signalling and indicators of oxidative stress in HAoSMCs. Furthermore, homoarginine reduced citrulline formation from arginine by nitric oxide (NO) synthase (NOS) isoforms. NO formation by NOS was reduced when using homoarginine as a substrate instead of arginine. The osteoinductive effects of homoarginine were mimicked by NOS inhibitor L-NAME and abolished by additional treatment with the NO donors DETA-NONOate and PAPA-NONOate or the antioxidants TEMPOL and TIRON. Furthermore, homoarginine augmented vascular calcification and aortic osteo-/chondrogenic signalling in mice after vitamin D3-overload, effects reversed by the NO donor molsidomine. CONCLUSION Homoarginine augments osteo-/chondrogenic transformation of VSMCs and vascular calcification, effects involving impaired NO formation from homoarginine.

[1]  F. Lang,et al.  Inhibition of Phosphate-Induced Vascular Smooth Muscle Cell Osteo-/Chondrogenic Signaling and Calcification by Bafilomycin A1 and Methylamine , 2015, Kidney and Blood Pressure Research.

[2]  W. März,et al.  Homoarginine and Clinical Outcomes in Renal Transplant Recipients: Results From the Assessment of Lescol in Renal Transplantation Study , 2015, Transplantation.

[3]  W. März,et al.  Homoarginine in the renal and cardiovascular systems , 2015, Amino Acids.

[4]  Y. Qu,et al.  Nitric oxide synthase in hypoxic or ischemic brain injury , 2015, Reviews in the neurosciences.

[5]  R. Wachter,et al.  Associations of methylarginines and homoarginine with diastolic dysfunction and cardiovascular risk factors in patients with preserved left ventricular ejection fraction. , 2014, Journal of cardiac failure.

[6]  C. Ayers,et al.  Homoarginine and Cardiovascular Outcome in the Population-Based Dallas Heart Study , 2014, Arteriosclerosis, thrombosis, and vascular biology.

[7]  C. Giachelli,et al.  A current understanding of vascular calcification in CKD. , 2014, American journal of physiology. Renal physiology.

[8]  N. Fujii,et al.  Dietary L-lysine prevents arterial calcification in adenine-induced uremic rats. , 2014, Journal of the American Society of Nephrology : JASN.

[9]  Jason L. Johnson.  Emerging regulators of vascular smooth muscle cell function in the development and progression of atherosclerosis. , 2014, Cardiovascular research.

[10]  D. Tsikas,et al.  GC–MS and GC–MS/MS measurement of the cardiovascular risk factor homoarginine in biological samples , 2014, Amino Acids.

[11]  K. Anseth,et al.  Aortic valve sclerosis in mice deficient in endothelial nitric oxide synthase. , 2014, American journal of physiology. Heart and circulatory physiology.

[12]  L. Juncos,et al.  Tempol blunts afferent arteriolar remodeling in chronic nitric oxide-deficient hypertension without normalizing blood pressure , 2014, Clinical and experimental hypertension.

[13]  W. März,et al.  Homoarginine, kidney function and cardiovascular mortality risk. , 2014, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[14]  B. Y. Erdoğan,et al.  Homoarginine, β-ODAP, and asparagine contents of grass pea landraces cultivated in Turkey. , 2014, Food chemistry.

[15]  S. Kato,et al.  Functional Cooperation between Vitamin D Receptor and Runx2 in Vitamin D-Induced Vascular Calcification , 2013, PloS one.

[16]  C. Gerloff,et al.  Homoarginine Levels Are Regulated by L-Arginine: Glycine Amidinotransferase and Affect Stroke Outcome Results From Human and Murine Studies , 2013, Circulation.

[17]  C. Zoccali,et al.  Homoarginine and Mortality in Pre-Dialysis Chronic Kidney Disease (CKD) Patients , 2013, PloS one.

[18]  J. Dekker,et al.  L-Homoarginine and L-arginine are antagonistically related to blood pressure in an elderly population: the Hoorn study , 2013, Journal of hypertension.

[19]  F. Kronenberg,et al.  Homoarginine and Progression of Chronic Kidney Disease: Results from the Mild to Moderate Kidney Disease Study , 2013, PloS one.

[20]  K. Rosenblatt,et al.  Spironolactone ameliorates PIT1-dependent vascular osteoinduction in klotho-hypomorphic mice. , 2013, The Journal of clinical investigation.

[21]  D. Carey,et al.  Novel Pathways in the Pathobiology of Human Abdominal Aortic Aneurysms , 2012, Pathobiology.

[22]  W. März,et al.  Homoarginine deficiency is associated with increased bone turnover , 2012, Osteoporosis International.

[23]  W. März,et al.  Homoarginine, heart failure, and sudden cardiac death in haemodialysis patients , 2011, European journal of heart failure.

[24]  Zhonghui Xu,et al.  Arterial injury promotes medial chondrogenesis in Sm22 knockout mice. , 2011, Cardiovascular research.

[25]  W. März,et al.  Low Serum Homoarginine Is a Novel Risk Factor for Fatal Strokes in Patients Undergoing Coronary Angiography , 2011, Stroke.

[26]  K. Rosenblatt,et al.  Hyperaldosteronism in Klotho-deficient mice. , 2010, American journal of physiology. Renal physiology.

[27]  W. März,et al.  Homoarginine, Cardiovascular Risk, and Mortality , 2010, Circulation.

[28]  H. Ryoo,et al.  Tumor necrosis factor-alpha increases alkaline phosphatase expression in vascular smooth muscle cells via MSX2 induction. , 2010, Biochemical and biophysical research communications.

[29]  R. Wadsworth.  Oxidative stress and the endothelium , 2008, Experimental physiology.

[30]  C. Lowenstein,et al.  Nitric oxide regulates vascular calcification by interfering with TGF- signalling. , 2008, Cardiovascular research.

[31]  J. Ribalta,et al.  Reference values for plasma concentrations of asymmetrical dimethylarginine (ADMA) and other arginine metabolites in men after validation of a chromatographic method. , 2007, Clinica chimica acta; international journal of clinical chemistry.

[32]  Su‐Li Cheng,et al.  Msx2 promotes cardiovascular calcification by activating paracrine Wnt signals. , 2005, The Journal of clinical investigation.

[33]  M. McKee,et al.  Smooth muscle cells deficient in osteopontin have enhanced susceptibility to calcification in vitro. , 2005, Cardiovascular research.

[34]  J. Passauer,et al.  Nitric oxide in chronic renal failure. , 2005, Kidney international.

[35]  Hyung-Suk Kim,et al.  L-Arginine uptake affects nitric oxide production and blood flow in the renal medulla. , 2004, American journal of physiology. Regulatory, integrative and comparative physiology.

[36]  K. Park,et al.  Nitric oxide-induced apoptosis is mediated by Bax/Bcl-2 gene expression, transition of cytochrome c, and activation of caspase-3 in rat vascular smooth muscle cells. , 2004, Clinica chimica acta; international journal of clinical chemistry.

[37]  M. Boy-Lefevre,et al.  Expression of collagen, osteocalcin, and bone alkaline phosphatase in a mineralizing rat osteoblastic cell culture , 1992, Calcified Tissue International.

[38]  T. Poulos,et al.  Structural Characterization and Kinetics of Nitric-oxide Synthase Inhibition by Novel N5-(Iminoalkyl)- and N5-(Iminoalkenyl)-ornithines* , 2003, Journal of Biological Chemistry.

[39]  G. London,et al.  Arterial media calcification in end-stage renal disease: impact on all-cause and cardiovascular mortality. , 2003, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[40]  M. Lerch,et al.  Vascular smooth muscle and nitric oxide synthase , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[41]  M. Wajner,et al.  In vitro stimulation of oxidative stress in cerebral cortex of rats by the guanidino compounds accumulating in hyperargininemia , 2001, Brain Research.

[42]  Y. Nabeshima,et al.  The progression of aging in klotho mutant mice can be modified by dietary phosphorus and zinc. , 2001, The Journal of nutrition.

[43]  H. Tsukahara,et al.  Increased oxidative stress in rats with chronic nitric oxide depletion:measurement of urinary 8-hydroxy-2′-deoxyguanosine excretion , 2000, Redox report : communications in free radical research.

[44]  J. Boucher,et al.  Substrate specificity of NO synthases: detailed comparison of L-arginine, homo-L-arginine, their N omega-hydroxy derivatives, and N omega-hydroxynor-L-arginine. , 1998, Biochemistry.

[45]  T. Hayakawa,et al.  Changes of serum alkaline phosphatase isoenzymes in fasted rats. , 1996, Journal of nutritional science and vitaminology.

[46]  E. Werner,et al.  Kinetics and Mechanism of Tetrahydrobiopterin-induced Oxidation of Nitric Oxide (*) , 1995, The Journal of Biological Chemistry.

[47]  Y. Yoshimura,et al.  Sensitivity of intestinal alkaline phosphatase to L-homoarginine and its regulation by subunit-subunit interaction. , 1994, Japanese journal of pharmacology.

[48]  A. Li,et al.  Arginine transport in human liver. Characterization and effects of nitric oxide synthase inhibitors. , 1993, Annals of Surgery.

[49]  S. Moncada,et al.  Identification of inhibitors of nitric oxide synthase that do not interact with the endothelial cell l‐arginine transporter , 1992, British journal of pharmacology.