Heart and Vessels

, Volume 29, Issue 5, pp 718–722 | Cite as

Reduction of inorganic phosphate-induced human smooth muscle cells calcification by inhibition of protein kinase A and p38 mitogen-activated protein kinase

  • Jeong-Hun KangEmail author
  • Riki Toita
  • Daisuke Asai
  • Tetsuji Yamaoka
  • Masaharu Murata
Short Communication


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.


Smooth muscle cell Calcification Protein kinase A Mitogen-activated protein kinase Alkaline phosphatase 



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.

Supplementary material

380_2013_427_MOESM1_ESM.pdf (214 kb)
Supplementary material 1 (PDF 213 kb)


  1. 1.
    Jono S, Shioi A, Ikari Y, Nishizawa Y (2006) Vascular calcification in chronic kidney disease. J Bone Miner Metab 24:176–181PubMedCrossRefGoogle Scholar
  2. 2.
    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 27:128–134PubMedCrossRefGoogle Scholar
  3. 3.
    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 28:12–18PubMedCrossRefGoogle Scholar
  4. 4.
    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 102:2636–2642PubMedCrossRefGoogle Scholar
  5. 5.
    Amann K (2008) Media calcification and intima calcification are distinct entities in chronic kidney disease. Clin J Am Soc Nephrol 3:1599–1605PubMedCrossRefGoogle Scholar
  6. 6.
    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–e17PubMedCrossRefGoogle Scholar
  7. 7.
    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 89:1147–1154PubMedCrossRefGoogle Scholar
  8. 8.
    Giachelli CM, Speer MY, Li X, Rajachar RM, Yang H (2005) Regulation of vascular calcification: roles of phosphate and osteopontin. Circ Res 96:717–722PubMedCrossRefGoogle Scholar
  9. 9.
    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 20:1254–1263PubMedCrossRefPubMedCentralGoogle Scholar
  10. 10.
    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 15:1392–1401PubMedCrossRefGoogle Scholar
  11. 11.
    Huang MS, Sage AP, Lu J, Demer LL, Tintut Y (2008) Phosphate and pyrophosphate mediated PKA-induced vascular cell calcification. Biochem Biophys Res Commun 374:553–558PubMedCrossRefPubMedCentralGoogle Scholar
  12. 12.
    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–C713PubMedCrossRefPubMedCentralGoogle Scholar
  13. 13.
    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 37:1531–1541PubMedCrossRefGoogle Scholar
  14. 14.
    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 583:1344–1348PubMedCrossRefPubMedCentralGoogle Scholar
  15. 15.
    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 46:572–580PubMedCrossRefGoogle Scholar
  16. 16.
    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 104:733–741CrossRefGoogle Scholar
  17. 17.
    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 34:525–530PubMedCrossRefGoogle Scholar
  18. 18.
    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:e33126PubMedCrossRefPubMedCentralGoogle Scholar
  19. 19.
    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:e17938PubMedCrossRefPubMedCentralGoogle Scholar
  20. 20.
    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 106:1807–1817PubMedCrossRefPubMedCentralGoogle Scholar

Copyright information

© Springer Japan 2013

Authors and Affiliations

  • Jeong-Hun Kang
    • 1
    Email author
  • Riki Toita
    • 2
  • Daisuke Asai
    • 3
  • Tetsuji Yamaoka
    • 1
  • Masaharu Murata
    • 4
  1. 1.Department of Biomedical EngineeringNational Cerebral and Cardiovascular Center Research InstituteSuitaJapan
  2. 2.Department of Biomaterials, Faculty of Dental ScienceKyushu UniversityFukuokaJapan
  3. 3.Department of MicrobiologySt. Marianna University School of MedicineKawasakiJapan
  4. 4.Department of Advanced Medical Initiatives, Faculty of Medical SciencesKyushu UniversityFukuokaJapan

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