title: NH4Cl Treatment Prevents Tissue Calcification in Klotho Deficiency.
abstract: Klotho, a cofactor in suppressing 1,25(OH)2D3 formation, is a powerful regulator of mineral metabolism. Klotho-hypomorphic mice (kl/kl) exhibit excessive plasma 1,25(OH)2D3, Ca(2+), and phosphate concentrations, severe tissue calcification, volume depletion with hyperaldosteronism, and early death. Calcification is paralleled by overexpression of osteoinductive transcription factor Runx2/Cbfa1, Alpl, and senescence-associated molecules Tgfb1, Pai-1, p21, and Glb1. Here, we show that NH4Cl treatment in drinking water (0.28 M) prevented soft tissue and vascular calcification and increased the life span of kl/kl mice >12-fold in males and >4-fold in females without significantly affecting extracellular pH or plasma concentrations of 1,25(OH)2D3, Ca(2+), and phosphate. NH4Cl treatment significantly decreased plasma aldosterone and antidiuretic hormone concentrations and reversed the increase of Runx2/Cbfa1, Alpl, Tgfb1, Pai-1, p21, and Glb1 expression in aorta of kl/kl mice. Similarly, in primary human aortic smooth muscle cells (HAoSMCs), NH4Cl treatment reduced phosphate-induced mRNA expression of RUNX2/CBFA1, ALPL, and senescence-associated molecules. In both kl/kl mice and phosphate-treated HAoSMCs, levels of osmosensitive transcription factor NFAT5 and NFAT5-downstream mediator SOX9 were higher than in controls and decreased after NH4Cl treatment. Overexpression of NFAT5 in HAoSMCs mimicked the effect of phosphate and abrogated the effect of NH4Cl on SOX9, RUNX2/CBFA1, and ALPL mRNA expression. TGFB1 treatment of HAoSMCs upregulated NFAT5 expression and prevented the decrease of phosphate-induced NFAT5 expression after NH4Cl treatment. In conclusion, NH4Cl treatment prevents tissue calcification, reduces vascular senescence, and extends survival of klotho-hypomorphic mice. The effects of NH4Cl on vascular osteoinduction involve decrease of TGFB1 and inhibition of NFAT5-dependent osteochondrogenic signaling.
authors: Departments of Physiology.
authors: Christina B
authors: CB
authors: Leibrock
authors: Departments of Physiology.
authors: Ioana
authors: I
authors: Alesutan
authors: Departments of Physiology.
authors: Jakob
authors: J
authors: Voelkl
authors: Departments of Physiology.
authors: Tatsiana
authors: T
authors: Pakladok
authors: Departments of Physiology.
authors: Diana
authors: D
authors: Michael
authors: Internal Medicine, and.
authors: Erwin
authors: E
authors: Schleicher
authors: Pathology, University of Tübingen, Tübingen, Germany; and.
authors: Zahra
authors: Z
authors: Kamyabi-Moghaddam
authors: Pathology, University of Tübingen, Tübingen, Germany; and.
authors: Leticia
authors: L
authors: Quintanilla-Martinez
authors: Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors: Makoto
authors: M
authors: Kuro-o
authors: Departments of Physiology, florian.lang@uni-tuebingen.de.
authors: Florian
authors: F
authors: Lang
mesh_terms: D000643:Ammonium Chloride / Q000627:therapeutic use*; D000818:Animals; D002114:Calcinosis / Q000209:etiology* / Q000517:prevention & control*; D005260:Female; D005966:Glucuronidase / Q000172:deficiency*; D000090265:Klotho Proteins; D008297:Male; D051379:Mice
chemical_list: D000643:Ammonium Chloride; D005966:Glucuronidase; D000090265:Klotho Proteins
keywords: CKD; aldosterone; mineral metabolism; vascular calcification
references: J Am Soc Nephrol. 1998 Dec;9(12 Suppl):S16-23
references: 11443763
references: Curr Opin Nephrol Hypertens. 2010 Jul;19(4):359-65
references: 20489614
references: J Am Soc Nephrol. 1994 Nov;5(5):1193-200
references: 7873729
references: Nephron. 2001 Feb;87(2):106-10
references: 11244303
references: Nephrol Dial Transplant. 2009 Aug;24(8):2488-96
references: 19329792
references: Pflugers Arch. 2009 May;458(1):39-52
references: 18758808
references: Stem Cell Rev. 2012 Sep;8(3):891-7
references: 22139789
references: Nephron Exp Nephrol. 2010;115(3):e60-8
references: 20424484
references: Can J Physiol Pharmacol. 2008 Sep;86(9):606-12
references: 18758509
references: Biochem J. 1993 Oct 1;295 ( Pt 1):11-4
references: 8216204
references: Curr Diab Rep. 2013 Jun;13(3):391-402
references: 23526400
references: J Am Soc Nephrol. 2013 Feb;24(2):179-89
references: 23138485
references: J Am Soc Nephrol. 2003 Sep;14(9 Suppl 4):S300-4
references: 12939385
references: Nephrol Dial Transplant. 2012 Jul;27(7):2650-7
references: 22802580
references: Am J Physiol Renal Physiol. 2013 Aug 1;305(3):F295-303
references: 23720342
references: FASEB J. 2010 Sep;24(9):3438-50
references: 20466874
references: Arterioscler Thromb Vasc Biol. 2011 Sep;31(9):2054-62
references: 21719763
references: Nature. 1997 Nov 6;390(6655):45-51
references: 9363890
references: Curr Opin Pharmacol. 2009 Apr;9(2):84-9
references: 19157979
references: Aging Cell. 2006 Apr;5(2):187-95
references: 16626397
references: Kidney Int. 2000 Sep;58(3):1023-32
references: 10972667
references: Kidney Int. 2008 Feb;73(4):407-14
references: 17989650
references: Kidney Blood Press Res. 2013;37(1):68-83
references: 23548827
references: Bone. 2013 Mar;53(1):94-102
references: 23219947
references: Am J Kidney Dis. 2014 Oct;64(4):542-9
references: 24953890
references: Circulation. 2005 Oct 25;112(17):2627-33
references: 16246962
references: J Am Soc Nephrol. 2014 Oct;25(10):2254-66
references: 24700866
references: J Am Soc Nephrol. 2010 Oct;21(10):1637-40
references: 20798261
references: Physiol Rev. 1981 Apr;61(2):296-434
references: 7012859
references: J Am Soc Nephrol. 2009 Jul;20(7):1453-64
references: 19478096
references: Biochim Biophys Acta. 2013 Mar;1830(3):2839-52
references: 23287548
references: Exp Biol Med (Maywood). 2012 Sep;237(9):1068-83
references: 22966145
references: J Am Soc Nephrol. 2015 Mar;26(3):515-23
references: 25150154
references: J Bone Miner Res. 2009 Nov;24(11):1847-55
references: 19419323
references: PLoS One. 2012;7(7):e39665
references: 22768306
references: Biol Chem. 2008 Mar;389(3):233-41
references: 18177265
references: Curr Opin Nephrol Hypertens. 2007 Jul;16(4):319-24
references: 17565273
references: Nat Rev Nephrol. 2013 Nov;9(11):650-60
references: 23774819
references: Pflugers Arch. 2010 Feb;459(3):499-508
references: 19841935
references: Am J Physiol Renal Physiol. 2010 Nov;299(5):F1171-7
references: 20719979
references: Curr Opin Nephrol Hypertens. 2013 Mar;22(2):193-7
references: 23380803
references: PLoS One. 2013;8(1):e55286
references: 23383138
references: Miner Electrolyte Metab. 1986;12(2):103-6
references: 3007964
references: Kidney Int. 2012 Dec;82(11):1184-94
references: 22832514
references: J Am Coll Cardiol. 2011 Apr 12;57(15):1634-5
references: 21474044
references: Circ Res. 2001 Dec 7;89(12):1147-54
references: 11739279
references: Hippokratia. 2009 Jul;13(3):139-40
references: 19918300
references: Kidney Int. 1998 Jun;53(6):1706-12
references: 9607203
references: Mech Ageing Dev. 2012 Nov-Dec;133(11-12):647-54
references: 23000105
references: Nephrol Dial Transplant. 2012 Aug;27(8):3056-62
references: 22851628
references: J Am Soc Nephrol. 2009 Nov;20(11):2371-9
references: 19713312
references: Circ Res. 2004 Sep 17;95(6):560-7
references: 15375022
references: Physiol Rev. 2007 Oct;87(4):1441-74
references: 17928589
references: Biogerontology. 2010 Feb;11(1):67-73
references: 19421891
references: J Bone Miner Res. 2010 May;25(5):1179-90
references: 19961337
references: Curr Aging Sci. 2011 Jul;4(2):128-36
references: 21235497
references: Kidney Int. 2008 Jan;73(2):192-9
references: 17978813
references: Adv Clin Chem. 2008;46:263-93
references: 19004192
references: Adv Chronic Kidney Dis. 2007 Jan;14(1):3-12
references: 17200038
references: J Am Soc Nephrol. 2011 Jan;22(1):124-36
references: 21115613
references: Nephrol Dial Transplant. 2003 Sep;18(9):1731-40
references: 12937218
references: Cell Physiol Biochem. 2010;26(6):1059-72
references: 21220937
references: Curr Opin Clin Nutr Metab Care. 2010 Jan;13(1):87-92
references: 19904201
references: Circ Res. 2012 Jul 6;111(2):245-59
references: 22773427
references: Arterioscler Thromb Vasc Biol. 2010 Sep;30(9):1741-50
references: 20522801
references: Am J Kidney Dis. 2014 Feb;63(2):311-7
references: 23932089
references: Cell Physiol Biochem. 2014;33(1):129-41
references: 24481040
references: Kidney Int. 2014 Mar;85(3):529-35
references: 24107852
references: Nephrol Dial Transplant. 2004 Aug;19(8):1986-92
references: 15173378
references: Biochem Biophys Res Commun. 2012 Jun 1;422(2):285-90
references: 22580282
references: Metab Brain Dis. 2010 Mar;25(1):31-8
references: 20336356
references: Neurochem Res. 2012 Nov;37(11):2379-87
references: 22544664
references: Curr Genomics. 2010 Dec;11(8):584-90
references: 21629436
references: J Clin Invest. 2013 Feb;123(2):812-22
references: 23298834
references: Am J Physiol. 1988 Dec;255(6 Pt 1):C844-56
references: 2849306
references: Science. 2005 Sep 16;309(5742):1829-33
references: 16123266
references: J Clin Invest. 1985 Aug;76(2):667-75
references: 2993363
references: Cardiovasc Res. 2008 Jan;77(1):221-30
references: 18006450
references: Tissue Eng Part A. 2011 Mar;17(5-6):597-606
references: 20874259
references: Clin J Am Soc Nephrol. 2010 Jan;5 Suppl 1:S23-30
references: 20089499
references: FASEB J. 2009 Feb;23(2):433-41
references: 18835926