Induction of bone-type alkaline phosphatase in human vascular smooth muscle cells: roles of tumor necrosis factor-alpha and oncostatin M derived from macrophages.
Inflammatory cells such as macrophages and T lymphocytes play an important role in vascular calcification associated with atherosclerosis and cardiac valvular disease. In particular, macrophages activated with cytokines derived from T lymphocytes such as interferon-gamma (IFN-gamma) may contribute to the development of vascular calcification. Moreover, we have shown the stimulatory effect of 1alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3) on in vitro calcification through increasing the expression of alkaline phosphatase (ALP), an ectoenzyme indispensable for bone mineralization, in vascular smooth muscle cells. Therefore, we hypothesized that macrophages may induce calcifying phenotype, especially the expression of ALP in human vascular smooth muscle cells (HVSMCs) in the presence of IFN-gamma and 1,25(OH)2D3. To test this hypothesis, we used cocultures of HVSMCs with human monocytic cell line (THP-1) or peripheral blood monocytes (PBMCs) in the presence of IFN-gamma and 1,25(OH)2D3. THP-1 cells or PBMCs induced ALP activity and its gene expression in HVSMCs and the cells with high expression of ALP calcified their extracellular matrix by the addition of beta-glycerophosphate. Thermostability and immunoassay showed that ALP induced in HVSMCs was bone-specific enzyme. We further identified tumor necrosis factor-alpha (TNF-alpha) and oncostatin M (OSM) as major factors inducing ALP in HVSMCs in the culture supernatants of THP-1 cells. TNF-alpha and OSM, only when applied together, increased ALP activities and in vitro calcification in HVSMCs in the presence of IFN-gamma and 1,25(OH)2D3. These results suggest that macrophages may contribute to the development of vascular calcification through producing various inflammatory mediators, especially TNF-alpha and OSM.
Department of Cardiovascular Medicine, Osaka City University Graduate School of Medicine, Osaka, Japan. as@msic.med.osaka-cu.ac.jp
Atsushi
A
Shioi
Miwako
M
Katagi
Yasuhisa
Y
Okuno
Katsuhito
K
Mori
Shuichi
S
Jono
Hidenori
H
Koyama
Yoshiki
Y
Nishizawa
D000469:Alkaline Phosphatase / Q000096:biosynthesis* / Q000235:genetics; D001842:Bone and Bones / Q000201:enzymology; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004790:Enzyme Induction / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D008264:Macrophages / Q000737:chemistry / Q000166:cytology; D009000:Monocytes / Q000737:chemistry / Q000166:cytology / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D053683:Oncostatin M; D053495:Osteopontin; D053244:Osteoprotegerin; D010455:Peptides / Q000494:pharmacology; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D018124:Receptors, Tumor Necrosis Factor; D012795:Sialoglycoproteins / Q000378:metabolism; D013250:Steroid Hydroxylases / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D017077:Culture Media, Conditioned; D006023:Glycoproteins; C508351:OSM protein, human; D053244:Osteoprotegerin; D010455:Peptides; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; C506030:TNFRSF11B protein, human; D014409:Tumor Necrosis Factor-alpha; D053495:Osteopontin; D053683:Oncostatin M; D007371:Interferon-gamma; D013250:Steroid Hydroxylases; C406322:vitamin D 1-alpha hydroxylase; D000469:Alkaline Phosphatase; D002118:Calcium