Induction of bone-type alkaline phosphatase in human vascular smooth muscle cells -: Roles of tumor necrosis factor-α and oncostatin M derived from macrophages
Induction of bone-type alkaline phosphatase in human vascular smooth muscle cells -: Roles of tumor necrosis factor-α and oncostatin M derived from macrophages
复制标题
DOI：
10.1161/01.res.0000026421.61398.f2
复制
发表时间：
2002-07-12
影响因子：
20.1
通讯作者：
Nishizawa, Y
中科院分区：
文献类型：
--
作者：
Shioi, A;Katagi, M;Nishizawa, Y
巨噬细胞和T淋巴细胞等炎症细胞在动脉粥样硬化和心脏瓣膜疾病相关的血管钙化中起重要作用。特别是，巨噬细胞被来自T淋巴细胞的细胞因子激活，如干扰素- γ (ifn - γ)，可能有助于血管钙化的发展。此外，我们已经证明了1 α，25-二羟基维生素D-3 (1,25（OH)(2)D-3）通过增加血管平滑肌细胞中骨矿化不可或缺的外酶碱性磷酸酶（ALP）的表达，对体外钙化的刺激作用。因此，我们假设巨噬细胞可能诱导钙化表型，特别是在ifn - γ和1,25(OH)(2)D-3存在的情况下，人血管平滑肌细胞（HVSMCs）中ALP的表达。为了验证这一假设，我们将HVSMCs与人单核细胞系（THP-1）或外周血单核细胞（PBMCs）在ifn - γ和1,25 (OH)(2)D-3，存在下共培养。THP-1细胞或PBMCs诱导HVSMCs中ALP活性及其基因表达，ALP高表达的细胞通过添加β -甘油磷酸酯使其细胞外基质钙化。热稳定性和免疫分析表明，在HVSMCs中诱导的ALP是骨特异性酶。我们进一步在THP-1细胞培养上清液中发现肿瘤坏死因子- α (tnf - α)和肿瘤抑制素M （OSM）是HVSMCs中诱导ALP的主要因子。tnf - α和OSM仅在同时应用时，在ifn - γ和1,25(OH)2D3存在的情况下，增加了HVSMCs的ALP活性和体外钙化。这些结果表明，巨噬细胞可能通过产生各种炎症介质，特别是tnf - α和OSM，促进血管钙化的发展。
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 D-3 (1,25(OH)(2)D-3) 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)(2)D-3. 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 l,25(OH)(2)D-3,. 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.