Abstract
High levels of serum phosphate are associated with calcification of human smooth muscle cells (HSMCs). We investigated whether inhibition of protein kinase A (PKA) and mitogen-activated protein kinase (MAPK) signals [p38, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK)] can reduce inorganic phosphate (Pi)-induced HSMC calcification. Inhibition of PKA or p38 MAPK by inhibitors or small interfering RNAs (siRNAs) reduced Ca levels and alkaline phosphatase activities in HSMCs treated with high Pi, but inhibition of ERK1/2 and JNK showed no significant changes. Moreover, there were no significant changes in cell viability on adding siRNAs and three inhibitors (PKA, p38, and MEK1/2), but JNK inhibitor slightly reduced cell viability. These results show that PKA and p38 MAPK are involved in the Pi-induced calcification of HSMCs, and may be good targets for reducing vascular calcification.
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References
Jono S, Shioi A, Ikari Y, Nishizawa Y (2006) Vascular calcification in chronic kidney disease. J Bone Miner Metab 24:176–181
Ueda H, Harimoto K, Tomoyama S, Tamaru H, Miyawaki M, Mitsusada N, Yasuga Y, Hiraoka H (2012) Relation of cardiovascular risk factors and angina status to obstructive coronary artery disease according to categorical coronary artery calcium score. Heart Vessels 27:128–134
Farrag A, Bakhoum S, Salem MA, El-Faramawy A, Gergis E (2013) The association between extracoronary calcification and coronary artery disease in patients with type 2 diabetes mellitus. Heart Vessels 28:12–18
Tintut Y, Patel J, Parhami F, Demer LL (2000) Tumor necrosis factor-α promotes in vitro calcification of vascular cells via the cAMP pathway. Circulation 102:2636–2642
Amann K (2008) Media calcification and intima calcification are distinct entities in chronic kidney disease. Clin J Am Soc Nephrol 3:1599–1605
Jono S, McKee MD, Murry CE, Shioi A, Nishizawa Y, Mori K, Morii H, Giachelli CM (2000) Phosphate regulation of vascular smooth muscle cell calcification. Circ Res 87:e10–e17
Steitz SA, Speer MY, Curinga G, Yang HY, Haynes P, Aebersold R, Schinke T, Karsenty G, Giachelli CM (2001) Smooth muscle cell phenotype transition associated with calcification: upregulation of Cbfa1 and downregulation of smooth muscle lineage markers. Circ Res 89:1147–1154
Giachelli CM, Speer MY, Li X, Rajachar RM, Yang H (2005) Regulation of vascular calcification: roles of phosphate and osteopontin. Circ Res 96:717–722
Zarjou A, Jeney V, Arosio P, Poli M, Antal-Szalmás P, Agarwal A, Balla G, Balla J (2009) Ferritin prevents calcification and osteoblastic differentiation of vascular smooth muscle cells. J Am Soc Nephrol 20:1254–1263
Lomashvili KA, Cobbs S, Hennigar RA, Hardcastle KI, O’Neill C (2004) Phosphate-induced vascular calcification: role of pyrophosphate and osteopontin. J Am Soc Nephrol 15:1392–1401
Huang MS, Sage AP, Lu J, Demer LL, Tintut Y (2008) Phosphate and pyrophosphate mediated PKA-induced vascular cell calcification. Biochem Biophys Res Commun 374:553–558
Prosdocimo DA, Wyler SC, Romani AM, O’Neill WC, Dubyak GR (2010) Regulation of vascular smooth muscle cell calcification by extracellular pyrophosphate homeostasis: synergistic modulation by cyclic AMP and hyperphosphatemia. Am J Physiol Cell Physiol 298:C702–C713
Simmons CA, Nikolovski J, Thornton AJ, Matlis S, Mooney DJ (2004) Mechanical stimulation and mitogen-activated protein kinase signaling independently regulate osteogenic differentiation and mineralization by calcifying vascular cells. J Biomech 37:1531–1541
Hsu JJ, Lu J, Huang MS, Geng Y, Sage AP, Bradley MN, Tontonoz P, Demer LL, Tintut Y (2009) T0901317, an LXR agonist, augments PKA-induced vascular cell calcification. FEBS Lett 583:1344–1348
Tanikawa T, Okada Y, Tanikawa R, Tanaka Y (2009) Advanced glycation end products induce calcification of vascular smooth muscle cells through RAGE/p38 MAPK. J Vasc Res 46:572–580
Speer MY, Yang HY, Brabb T, Leaf E, Look A, Lin WL, Frutkin A, Dichek D, Giachelli CM (2009) Smooth muscle cells give rise to osteochondrogenic precursors and chondrocytes in calcifying arteries. Cir Res 104:733–741
Liao XB, Zhou XM, Li JM, Yang JF, Tan ZP, Hu ZW, Liu W, Lu Y, Yuan LQ (2008) Taurine inhibits osteoblastic differentiation of vascular smooth muscle cells via the ERK pathway. Amino Acids 34:525–530
Liang QH, Jiang Y, Zhu X, Cui RR, Liu GY, Liu Y, Wu SS, Liao ZB, Xie H, Zhou HD, Wu XP, Yuan LQ, Liao EY (2012) Ghrelin attenuates the osteoblastic differentiation of vascular smooth muscle cells through the ERK pathway. PLoS One 7:e33126
Shan PF, Lu Y, Cui RR, Jiang Y, Yuan LQ, Liao EY (2011) Apelin attenuates the osteoblastic differentiation of vascular smooth muscle cells. PLoS One 6:e17938
Miyazaki-Anzai S, Levi M, Kratzer A, Ting TC, Lewis LB, Miyazaki M (2010) Farnesoid X receptor activation prevents the development of vascular calcification in ApoE−/− mice with chronic kidney disease. Circ Res 106:1807–1817
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This work was financially supported by a grant-in-aid for Scientific Research (B) (KAKENHI Grant Number 23310085) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan.
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Kang, JH., Toita, R., Asai, D. et al. Reduction of inorganic phosphate-induced human smooth muscle cells calcification by inhibition of protein kinase A and p38 mitogen-activated protein kinase. Heart Vessels 29, 718–722 (2014). https://doi.org/10.1007/s00380-013-0427-x
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DOI: https://doi.org/10.1007/s00380-013-0427-x