caa a22 4500 606236104 CHVBK 20210128101246.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s11010-015-2432-0 doi (NATIONALLICENCE)springer-10.1007/s11010-015-2432-0 Vascular smooth muscle cell differentiation to an osteogenic phenotype involves matrix metalloproteinase-2 modulation by homocysteine [Elektronische Daten] [Tingjiao Liu, Jinghan Lin, Ting Ju, Lei Chu, Liming Zhang] Arterial calcification is common in vascular diseases and involves conversion of vascular smooth muscle cells (VSMCs) to an osteoblast phenotype. Clinical studies suggest that the development of atherosclerosis can be promoted by homocysteine (HCY), but the mechanisms remain unclear. Here, we determined whether increases in HCY levels lead to an increase in VSMC calcification and differentiation, and examined the role of an extracellular matrix remodeler, matrix metalloproteinase-2 (MMP-2). Rat VSMCs were exposed to calcification medium in the absence or presence of HCY (10, 100 or 200μmol/L) or an MMP-2 inhibitor (10−6 or 10−5 mol/L). MTT assays were performed to determine the cytotoxicity of the MMP-2 inhibitor in calcification medium containing 200μmol/L HCY. Calcification was assessed by measurements of calcium deposition and alkaline phosphatase (ALP) activity as well as von Kossa staining. Expression of osteocalcin, bone morphogenetic protein (BMP)-2, and osteopontin, and MMP-2 was determined by immunoblotting. Calcification medium induced osteogenic differentiation of VSMCs. HCY promoted calcification, increased osteocalcin and BMP-2 expression, and decreased expression of osteopontin. MMP-2 expression was increased by HCY in a dose-dependent manner in VSMCs exposed to both control and calcification medium. The MMP-2 inhibitor decreased the calcium content and ALP activity, and attenuated the osteoblastic phenotype of VSMCs. Vascular calcification and osteogenic differentiation of VSMCs were positively regulated by HCY through increased/restored MMP-2 expression, increased expression of calcification proteins, and decreased anti-calcification protein levels. In summary, MMP-2 inhibition may be a protective strategy against VSMC calcification. Springer Science+Business Media New York, 2015 Calcification nationallicence Homocysteine nationallicence Matrix metalloproteinase-2 nationallicence Osteogenic phenotype nationallicence Liu Tingjiao Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China aut Lin Jinghan Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China aut Ju Ting Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China aut Chu Lei Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China aut Zhang Liming Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 406/1-2(2015-08-01), 139-149 0300-8177 406:1-2<139 2015 406 11010 https://doi.org/10.1007/s11010-015-2432-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-015-2432-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Liu Tingjiao Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China aut NATIONALLICENCE 700 1- Lin Jinghan Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China aut NATIONALLICENCE 700 1- Ju Ting Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China aut NATIONALLICENCE 700 1- Chu Lei Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China aut NATIONALLICENCE 700 1- Zhang Liming Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 406/1-2(2015-08-01), 139-149 0300-8177 406:1-2<139 2015 406 11010