Similar articles

Abstract: In this paper, we investigate the effect and the possible mechanism of high glucose levels on the calcification of human aortic smooth muscle cells (HASMCs). HASMCs were divided into four groups: normal glucose group (NG), osmolality control group (OC), high glucose group (HG, HASMCs culture medium containing 30 mmol/L glucose), and high glucose plus recombinant human noggin protein (bone morphogenetic protein-2 (BMP-2) antagonist) group (HN). The mRNA levels and the protein expressions of BMP-2 and core binding factor alpha-1 (Cbfα;-1) were measured by real-time quantitative polymerase chain reaction (PCR) and Western blot. After induced by 10 mmol/L β-glycerol phosphoric acid, cells were harvested for assessments of alkaline phosphatase (ALP) activities at Days 1, 2, and 3, and intracellular calcium contents at Days 7 and 14, respectively. High glucose levels increased the mRNA levels and the protein expressions of BMP-2 and Cbfα-1 (P<0.05). The expression of Cbfα-1 was partially blocked by noggin protein (P<0.05), while BMP-2 was not (P>0.05). After being induced by β-glycerol phosphoric acid, high glucose levels increased the ALP activity [(48.63±1.03) vs. (41.42±2.28) U/mg protein, Day 3; P<0.05] and the intracellular calcium content [(2.76±0.09) vs. (1.75±0.07) μmol/mg protein, Day 14; P<0.05] in a time-dependent manner when compared with the NG group, while the ALP activity could not be blocked by noggin protein [(48.63±1.03) vs. (47.37±0.97) U/mg protein, Day 3; P>0.05]. These results show that high glucose levels can evoke the calcification of HASMCs by inducing osteoblastic trans-differentiation and intracellular calcium deposition via the BMP-2/Cbfα-1 pathway, which can be partially blocked by noggin protein.

[1]Attisano, L., Wrana, J.L., 2002. Signal transduction by the TGF-β superfamily. Science, 296(5573):1646-1647.

[2]Busch, C., Drews, U., Eisele, S.R., Garbe, C., Oppitz, M., 2008. Noggin blocks invasive growth of murine B16-F1 melanoma cells in the optic cup of the chick embryo. Int. J. Cancer, 122(3):526-533.

[3]Canalis, E., Economides, A.N., Gazzerro, E., 2003. Bone morphogenetic proteins, their antagonists, and the skeleton. Endocr. Rev., 24(2):218-235.

[4]Chen, N.X., O′Neill, K.D., Duan, D., Moe, S.M., 2002. Phosphorus and uremic serum up-regulate osteopontin expression in vascular smooth muscle cells. Kidney Int., 62(5):1724-1731.

[5]Chen, N.X., Duan, D., O′Neill, K.D., Moe, S.M., 2006. High glucose increases the expression of Cbfα1 and BMP-2 and enhances the calcification of vascular smooth muscle cells. Nephrol. Dial. Transplant., 21(12):3435-3442.

[6]Conley, B.A., Smith, J.D., Guerrero-Esteo, M., Bernabeu, C., Vary, C.P., 2000. Endoglin, a TGF-β receptor-associated protein, is expressed by smooth muscle cells in human atherosclerotic plaques. Atherosclerosis, 153(2):323-335.

[7]Dhore, C.R., Cleutjens, J.P., Lutgens, E., Cleutjens, K.B., Geusens, P.P., Kitslaar, P.J., Tordoir, J.H., Spronk, H.M., Vermeer, C., Daemen, M.J., 2001. Differential expression of bone matrix regulatory proteins in human atherosclerotic plaques. Arterioscler. Thromb. Vasc. Biol., 21(12):1998-2003.

[8]Doherty, T.M., Fitzpatrick, L.A., Inoue, D., Qiao, J.H., Fishbein, M.C., Detrano, R.C., Shah, P.K., Rajavashisth, T.B., 2004. Molecular, endocrine, and genetic mechanisms of arterial calcification. Endocr. Rev., 25(4):629-672.

[9]Lee, K.S., Kim, H.J., Li, Q.L., Chi, X.Z., Ueta, C., Komori, T., Wozney, J.M., Kim, E.G., Choi, J.Y., Ryoo, H.M., et al., 2000. Runx2 is a common target of transforming growth factor-1 and bone morphogenetic protein 2, and cooperation between Runx2 and Smad5 induces osteoblast-specific gene expression in the pluripotent mesenchymal precursor cell line C2C12. Mol. Cell. Biol., 20(23):8783-8792.

[10]Lehto, S., Niskanen, L., Suhonen, M., Rönnemaa, T., Laakso, M., 1996. Medial artery calcification a neglected harbinger of cardiovascular complications in non-insulin-dependent diabetes mellitus. Arterioscler. Thromb. Vasc. Biol., 16(8):978-983.

[11]Moe, S.M., Chen, N.X., 2004. Pathophysiology of vascular calcification in chronic kidney disease. Circ. Res., 95(6):560-567.

[12]Moe, S.M., Duan, D., Doehle, B.P., O′Neill, K.D., Chen, N.X., 2003. Uremia induces the osteoblast differentiation factor Cbfα1 in human blood vessels. Kidney Int., 63(3):1003-1011.

[13]Mori, K., Shioi, A., Jono, S., Nishizawa, Y., Morii, H., 1999. Dexamethasone enhances in vitro vascular calcification by promoting osteoblastic differentiation of vascular smooth muscle cells. Arterioscler. Thromb. Vasc. Biol., 19(9):2112-2118.

[14]Otto, F., Lubbert, M., Stock, M., 2003. Upstream and downstream targets of RUNX proteins. J. Cell Biochem., 89(1):9-18.

[15]Parhami, F., Basseri, B., Hwang, J., Tintut, Y., Demer, L.L., 2002. High density lipoprotein regulates calcification of vascular cells. Circ. Res., 91(7):570-576.

[16]Shanahan, C.M., Cary, N.R., Metcalfe, J.C., Weissberg, P.L., 1994. High expression of genes for calcification-regulating proteins in human atherosclerotic plaques. J. Clin. Invest., 93(6):2393-2402.

[17]Shioi, A., Nishizawa, Y., Jono, S., Koyama, H., Hosoi, M., Morii, H., 1995. β-glycerophosphate accelerates calcification in cultured bovine vascular smooth muscle cells. Arterioscler. Thromb. Vasc. Biol., 15(11):2003-2009.

[18]Shioi, A., Katagi, M., Okuno, Y., Mori, K., Jono, S., Koyama, H., Nishizawa, Y., 2002. Induction of bone-type alkaline phosphatase in human vascular smooth muscle cells: roles of tumor necrosis factor-α and oncostatin M derived from macrophages. Circ. Res., 91(1):9-16.

[19]Steitz, S.A., Speer, M.Y., Curinga, G., Yang, H.Y., Haynes, P., Aebersold, R., Schinke, T., Karsenty, G., Giachelli, C.M., 2001. Smooth muscle cell phenotypic transition associated with calcification: upregulation of Cbfα1 and downregulation of smooth muscle lineage markers. Circ. Res., 89(12):1147-1154.

[20]Takayama, K., Suzuki, A., Manaka, T., Taguchi, S., Hashimoto, Y., Imai, Y., Wakitani, S., Takaoka, K., 2009. RNA interference for Noggin enhances the biological activity of bone morphogenetic proteins in vivo and in vitro. J. Bone Miner. Metab., 27(4):402-411.

[21]Tintut, Y., Patel, J., Parhami, F., 2000. Tumor necrosis factor-α promotes in vitro calcification of vascular cells via the cAMP pathway. Circulation, 102(21):2636-2642.

[22]Yuasa, S., Fukuda, K., 2008. Multiple roles for BMP signaling in cardiac development. Drug Discov. Today: Ther. Strategies, 5(4):209-214.

[23]Zhu, W., Kim, J., Cheng, C., Rawlins, B.A., Boachie-Adjei, O., Crystal, R.G., Hidaka, C., 2006. Noggin regulation of bone morphogenesis protein (BMP) 2/7 heterodimer activity in vitro. Bone, 39(1):61-71.