Fibronectin enhances in vitro vascular calcification by promoting osteoblastic differentiation of vascular smooth muscle cells via ERK pathway.
The process of vascular calcification presents several features similar to osteogenesis in which fibronectin (FN) acts as a regulator of osteoblastic differentiation and the ERK signal pathway is involved. In order to find whether FN promotes the osteoblastic differentiation of vascular smooth muscle cells (VSMCs) through the ERK signal pathway, we investigated the effect of FN on the calcification of VSMCs by using an in vitro cell model. VSMCs cultured in plates with FN (0-20 microg/cm2) coating were induced to calcify by 10 mM sodium beta-glycerophosphate (beta-GP). FN exacerbated VSMC calcification in a dose- and time-dependent manner, as indicated by the number of calcifying nodules per slide and the amount of calcium in the deposition. Data from RT-PCR and immunoblotting assay revealed that FN also enhanced the expression of several phenotypic markers of osteoblasts, including alkaline phosphatase (ALP) activity, osteocalcin (OC), and Osf2/Cbfa1, a key transcription factor in osteoblastic differentiation. Furthermore, a specific inhibitor for ERK, PD98059 (10 microM), significantly suppressed the effect of FN on calcification and phenotypic marker expression. These findings seem to suggest that FN enhanced vascular calcification by promoting the osteoblastic differentiation of VSMCs via ERK signal pathway.
Ding
Hai-Tao
HT
Department of Chemical Biology, Peking University School of Pharmaceutical Sciences, 38 Xueyuan Road, Beijing 100083, People's Republic of China.
Wang
Chen-Guang
CG
Zhang
Tian-Lan
TL
Wang
Kui
K
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002114:Calcinosis; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism*; D005419:Flavonoids / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000502:physiology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000502:physiology; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology*; D051381:Rats
D015415:Biomarkers; D004791:Enzyme Inhibitors; D005353:Fibronectins; D005419:Flavonoids; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one