Molecular and Cellular Biochemistry

, Volume 420, Issue 1–2, pp 151–160 | Cite as

Curcumin attenuates osteogenic differentiation and calcification of rat vascular smooth muscle cells

  • Menglin Hou
  • Yan Song
  • Zhenlin Li
  • Chufan Luo
  • Jing-Song Ou
  • Huimin Yu
  • Jianyun Yan
  • Lihe Lu


Vascular calcification has been considered as a biological process resembling bone formation involving osteogenic differentiation. It is a major risk factor for cardiovascular morbidity and mortality. Previous studies have shown the protective effects of curcumin on cardiovascular diseases. However, whether curcumin has effects on osteogenic differentiation and calcification of vascular smooth muscle cells (VSMCs) has not been reported. In the present study, we used an in vitro model of VSMC calcification to investigate the role of curcumin in the progression of rat VSMC calcification. Curcumin treatment significantly reduced calcification of VSMCs in a dose-dependent manner, detected by alizarin red staining and calcium content assay. Similarly, ALP activity and expression of bone-related molecules including Runx2, BMP2, and Osterix were also decreased in VSMCs treated with curcumin. In addition, flow cytometry analysis and caspase-3 activity assay revealed that curcumin treatment significantly suppressed apoptosis of VSMCs, which plays an important role during vascular calcification. Furthermore, we found that pro-apoptotic molecules including p-JNK and Bax were up-regulated in VSMCs treated with calcifying medium, but they were reduced in VSMCs after curcumin treatment. However, curcumin treatment has no effect on expression of NF-κB p65. Taken together, these findings suggest that curcumin attenuates apoptosis and calcification of VSMCs, presumably via inhibition of JNK/Bax signaling pathway.


Curcumin Vascular smooth muscle cells Calcification Apoptosis 



This work was supported by the National Natural Science Foundation of China (81470488 to J.Y.Y, 81325001 to J.S.O), and the Guangdong Natural Science Foundation, China (2015A030313260 to J.Y.Y, 2015A030312009 to J.S.O, 2016A030313226 to L.H.L), the Science and Technology Planning Project of Guangdong Province (2014A020221084 to L.H.L), and the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry (to J.Y.Y).

Compliance with ethical standards

Conflict of interest

All authors have declared no conflicts of interest.


  1. 1.
    Shanahan CM, Crouthamel MH, Kapustin A, Giachelli CM (2011) Arterial calcification in chronic kidney disease: key roles for calcium and phosphate. Circ Res 109:697–711CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    London GM, Guerin AP, Marchais SJ, Metivier F, Pannier B, Adda H (2003) Arterial media calcification in end-stage renal disease: impact on all-cause and cardiovascular mortality. Nephrol Dial Transplant 18:1731–1740CrossRefPubMedGoogle Scholar
  3. 3.
    Wilson PW, Kauppila LI, O’Donnell CJ, Kiel DP, Hannan M, Polak JM, Cupples LA (2001) Abdominal aortic calcific deposits are an important predictor of vascular morbidity and mortality. Circulation 103:1529–1534CrossRefPubMedGoogle Scholar
  4. 4.
    Lanzer P, Boehm M, Sorribas V, Thiriet M, Janzen J, Zeller T, St Hilaire C, Shanahan C (2014) Medial vascular calcification revisited: review and perspectives. Eur Heart J 35:1515–1525CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Evrard S, Delanaye P, Kamel S, Cristol JP, Cavalier E (2015) Vascular calcification: from pathophysiology to biomarkers. Clin Chim Acta 438:401–414CrossRefPubMedGoogle Scholar
  6. 6.
    Sage AP, Tintut Y, Demer LL (2010) Regulatory mechanisms in vascular calcification. Nat Rev Cardiol 7:528–536CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Shanahan CM, Cary NR, Salisbury JR, Proudfoot D, Weissberg PL, Edmonds ME (1999) Medial localization of mineralization-regulating proteins in association with Monckeberg’s sclerosis: evidence for smooth muscle cell-mediated vascular calcification. Circulation 100:2168–2176CrossRefPubMedGoogle Scholar
  8. 8.
    Li X, Yang HY, Giachelli CM (2008) BMP-2 promotes phosphate uptake, phenotypic modulation, and calcification of human vascular smooth muscle cells. Atherosclerosis 199:271–277CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Steitz SA, Speer MY, Curinga G, Yang HY, Haynes P, Aebersold R, Schinke T, Karsenty G, Giachelli CM (2001) Smooth muscle cell phenotypic transition associated with calcification: upregulation of Cbfa1 and downregulation of smooth muscle lineage markers. Circ Res 89:1147–1154CrossRefPubMedGoogle Scholar
  10. 10.
    Shroff R, Long DA, Shanahan C (2013) Mechanistic insights into vascular calcification in CKD. J Am Soc Nephrol 24:179–189CrossRefPubMedGoogle Scholar
  11. 11.
    Proudfoot D, Skepper JN, Hegyi L, Bennett MR, Shanahan CM, Weissberg PL (2000) Apoptosis regulates human vascular calcification in vitro: evidence for initiation of vascular calcification by apoptotic bodies. Circ Res 87:1055–1062CrossRefPubMedGoogle Scholar
  12. 12.
    Yan J, Stringer SE, Hamilton A, Charlton-Menys V, Gotting C, Muller B, Aeschlimann D, Alexander MY (2011) Decorin GAG synthesis and TGF-beta signaling mediate Ox-LDL-induced mineralization of human vascular smooth muscle cells. Arterioscler Thromb Vasc Biol 31:608–615CrossRefPubMedGoogle Scholar
  13. 13.
    Liao L, Zhou Q, Song Y, Wu W, Yu H, Wang S, Chen Y, Ye M, Lu L (2013) Ceramide mediates Ox-LDL-induced human vascular smooth muscle cell calcification via p38 mitogen-activated protein kinase signaling. PLoS One 8:e82379CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Epstein J, Sanderson IR, Macdonald TT (2010) Curcumin as a therapeutic agent: the evidence from in vitro, animal and human studies. Br J Nutr 103:1545–1557CrossRefPubMedGoogle Scholar
  15. 15.
    Yang X, Thomas DP, Zhang X, Culver BW, Alexander BM, Murdoch WJ, Rao MN, Tulis DA, Ren J, Sreejayan N (2006) Curcumin inhibits platelet-derived growth factor-stimulated vascular smooth muscle cell function and injury-induced neointima formation. Arterioscler Thromb Vasc Biol 26:85–90CrossRefPubMedGoogle Scholar
  16. 16.
    Hasan ST, Zingg JM, Kwan P, Noble T, Smith D, Meydani M (2014) Curcumin modulation of high fat diet-induced atherosclerosis and steatohepatosis in LDL receptor deficient mice. Atherosclerosis 232:40–51CrossRefPubMedGoogle Scholar
  17. 17.
    Huang HC, Jan TR, Yeh SF (1992) Inhibitory effect of curcumin, an anti-inflammatory agent, on vascular smooth muscle cell proliferation. Eur J Pharmacol 221:381–384CrossRefPubMedGoogle Scholar
  18. 18.
    Yu YM, Lin HC (2010) Curcumin prevents human aortic smooth muscle cells migration by inhibiting of MMP-9 expression. Nutr Metab Cardiovasc Dis 20:125–132CrossRefPubMedGoogle Scholar
  19. 19.
    Meng Z, Yan C, Deng Q, Gao DF, Niu XL (2013) Curcumin inhibits LPS-induced inflammation in rat vascular smooth muscle cells in vitro via ROS-relative TLR4-MAPK/NF-kappaB pathways. Acta Pharmacol Sin 34:901–911CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Chen FY, Zhou J, Guo N, Ma WG, Huang X, Wang H, Yuan ZY (2015) Curcumin retunes cholesterol transport homeostasis and inflammation response in M1 macrophage to prevent atherosclerosis. Biochem Biophys Res Commun 467:872–878CrossRefPubMedGoogle Scholar
  21. 21.
    Hao F, Kang J, Cao Y, Fan S, Yang H, An Y, Pan Y, Tie L, Li X (2015) Curcumin attenuates palmitate-induced apoptosis in MIN6 pancreatic beta-cells through PI3 K/Akt/FoxO1 and mitochondrial survival pathways. Apoptosis 20:1420–1432CrossRefPubMedGoogle Scholar
  22. 22.
    Shroff RC, McNair R, Figg N, Skepper JN, Schurgers L, Gupta A, Hiorns M, Donald AE, Deanfield J, Rees L, Shanahan CM (2008) Dialysis accelerates medial vascular calcification in part by triggering smooth muscle cell apoptosis. Circulation 118:1748–1757CrossRefPubMedGoogle Scholar
  23. 23.
    Notoya M, Nishimura H, Woo JT, Nagai K, Ishihara Y, Hagiwara H (2006) Curcumin inhibits the proliferation and mineralization of cultured osteoblasts. Eur J Pharmacol 534:55–62CrossRefPubMedGoogle Scholar
  24. 24.
    Lewinska A, Wnuk M, Grabowska W, Zabek T, Semik E, Sikora E, Bielak-Zmijewska A (2015) Curcumin induces oxidation-dependent cell cycle arrest mediated by SIRT7 inhibition of rDNA transcription in human aortic smooth muscle cells. Toxicol Lett 233:227–238CrossRefPubMedGoogle Scholar
  25. 25.
    Chen Z, Xue J, Shen T, Ba G, Yu D, Fu Q (2016) Curcumin alleviates glucocorticoid-induced osteoporosis by protecting osteoblasts from apoptosis in vivo and in vitro. Clin Exp Pharmacol Physiol 43:268–276CrossRefPubMedGoogle Scholar
  26. 26.
    Dhanasekaran DN, Reddy EP (2008) JNK signaling in apoptosis. Oncogene 27:6245–6251CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Chen YR, Tan TH (1998) Inhibition of the c-Jun N-terminal kinase (JNK) signaling pathway by curcumin. Oncogene 17:173–178CrossRefPubMedGoogle Scholar
  28. 28.
    Papadakis ES, Finegan KG, Wang X, Robinson AC, Guo C, Kayahara M, Tournier C (2006) The regulation of Bax by c-Jun N-terminal protein kinase (JNK) is a prerequisite to the mitochondrial-induced apoptotic pathway. FEBS Lett 580:1320–1326CrossRefPubMedGoogle Scholar
  29. 29.
    Oltvai ZN, Milliman CL, Korsmeyer SJ (1993) Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74:609–619CrossRefPubMedGoogle Scholar
  30. 30.
    Reynolds JL, Joannides AJ, Skepper JN, McNair R, Schurgers LJ, Proudfoot D, Jahnen-Dechent W, Weissberg PL, Shanahan CM (2004) Human vascular smooth muscle cells undergo vesicle-mediated calcification in response to changes in extracellular calcium and phosphate concentrations: a potential mechanism for accelerated vascular calcification in ESRD. J Am Soc Nephrol 15:2857–2867CrossRefPubMedGoogle Scholar
  31. 31.
    Shroff RC, McNair R, Skepper JN, Figg N, Schurgers LJ, Deanfield J, Rees L, Shanahan CM (2010) Chronic mineral dysregulation promotes vascular smooth muscle cell adaptation and extracellular matrix calcification. J Am Soc Nephrol 21:103–112CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Son BK, Kozaki K, Iijima K, Eto M, Kojima T, Ota H, Senda Y, Maemura K, Nakano T, Akishita M, Ouchi Y (2006) Statins protect human aortic smooth muscle cells from inorganic phosphate-induced calcification by restoring Gas6-Axl survival pathway. Circ Res 98:1024–1031CrossRefPubMedGoogle Scholar
  33. 33.
    Yang H, Curinga G, Giachelli CM (2004) Elevated extracellular calcium levels induce smooth muscle cell matrix mineralization in vitro. Kidney Int 66:2293–2299CrossRefPubMedGoogle Scholar
  34. 34.
    Zhang L, Zhou M, Wang Y, Huang W, Qin G, Weintraub NL, Tang Y (2014) miR-92a inhibits vascular smooth muscle cell apoptosis: role of the MKK4-JNK pathway. Apoptosis 19:975–983CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Han Y, Wu G, Deng J, Tao J, Guo L, Tian X, Kang J, Zhang X, Yan C (2010) Cellular repressor of E1A-stimulated genes inhibits human vascular smooth muscle cell apoptosis via blocking P38/JNK MAP kinase activation. J Mol Cell Cardiol 48:1225–1235CrossRefPubMedGoogle Scholar
  36. 36.
    Zhang X, Zhou C, Zha X, Xu Z, Li L, Liu Y, Xu L, Cui L, Xu D, Zhu B (2015) Apigenin promotes osteogenic differentiation of human mesenchymal stem cells through JNK and p38 MAPK pathways. Mol Cell Biochem 407:41–50CrossRefPubMedGoogle Scholar
  37. 37.
    Wang B, Li F, Zhang C, Wei G, Liao P, Dong N (2016) High-mobility group box-1 protein induces osteogenic phenotype changes in aortic valve interstitial cells. J Thorac Cardiovasc Surg 151:255–262CrossRefPubMedGoogle Scholar
  38. 38.
    Suzuki A, Ghayor C, Guicheux J, Magne D, Quillard S, Kakita A, Ono Y, Miura Y, Oiso Y, Itoh M, Caverzasio J (2006) Enhanced expression of the inorganic phosphate transporter Pit-1 is involved in BMP-2-induced matrix mineralization in osteoblast-like cells. J Bone Miner Res 21:674–683CrossRefPubMedGoogle Scholar
  39. 39.
    Wu Y, Han X, Wang L, Diao Z, Liu W (2016) Indoxyl sulfate promotes vascular smooth muscle cell calcification via the JNK/Pit-1 pathway. Ren Fail 27:1–9CrossRefGoogle Scholar
  40. 40.
    Tsuruta F, Sunayama J, Mori Y, Hattori S, Shimizu S, Tsujimoto Y, Yoshioka K, Masuyama N, Gotoh Y (2004) JNK promotes Bax translocation to mitochondria through phosphorylation of 14-3-3 proteins. EMBO J 23:1889–1899CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Zhao MM, Xu MJ, Cai Y, Zhao G, Guan Y, Kong W, Tang C, Wang X (2011) Mitochondrial reactive oxygen species promote p65 nuclear translocation mediating high-phosphate-induced vascular calcification in vitro and in vivo. Kidney Int 79:1071–1079CrossRefPubMedGoogle Scholar
  42. 42.
    Zhao G, Xu MJ, Zhao MM, Dai XY, Kong W, Wilson GM, Guan Y, Wang CY, Wang X (2012) Activation of nuclear factor-kappa B accelerates vascular calcification by inhibiting ankylosis protein homolog expression. Kidney Int 82:34–44CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Wongcharoen W, Phrommintikul A (2009) The protective role of curcumin in cardiovascular diseases. Int J Cardiol 133:145–151CrossRefPubMedGoogle Scholar
  44. 44.
    Pan Y, Wang Y, Cai L, Cai Y, Hu J, Yu C, Li J, Feng Z, Yang S, Li X, Liang G (2012) Inhibition of high glucose-induced inflammatory response and macrophage infiltration by a novel curcumin derivative prevents renal injury in diabetic rats. Br J Pharmacol 166:1169–1182CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Pan Y, Wang Y, Zhao Y, Peng K, Li W, Wang Y, Zhang J, Zhou S, Liu Q, Li X, Cai L, Liang G (2014) Inhibition of JNK phosphorylation by a novel curcumin analog prevents high glucose-induced inflammation and apoptosis in cardiomyocytes and the development of diabetic cardiomyopathy. Diabetes 63:3497–3511CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Pathophysiolgy, Zhongshan Medical SchoolSun Yat-Sen UniversityGuangzhouChina
  2. 2.Department of Histology and Embryology, Key Laboratory of Tissue Construction and Detection of Guangdong ProvinceSouthern Medical UniversityGuangzhouChina
  3. 3.Department of Cardiology, The First Affiliated HospitalSun Yat-Sen UniversityGuangzhouChina
  4. 4.Division of Cardiac Surgery, The First Affiliated HospitalSun Yat-Sen UniversityGuangzhouChina
  5. 5.The Key Laboratory of Assisted Circulation, Ministry of HealthSun Yat-Sen UniversityGuangzhouChina
  6. 6.Department of Cardiology, Guangdong General HospitalGuangdong Academy of Medical Sciences and Guangdong Cardiovascular InstituteGuangzhouChina

Personalised recommendations