Osteogenic responses in fibroblasts activated by elastin degradation products and transforming growth factor-β1 -: Role of myofibroblasts in vascular calcification
Osteogenic responses in fibroblasts activated by elastin degradation products and transforming growth factor-β1 -: Role of myofibroblasts in vascular calcification
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DOI：
10.2353/ajpath.2007.060930
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发表时间：
2007-07-01
影响因子：
6
通讯作者：
Vyavahare, Narendra R.
中科院分区：
文献类型：
--
作者：
Simionescu, Agneta;Sirnionescu, Dan T.;Vyavahare, Narendra R.
我们的目标是确定成纤维细胞在内侧血管钙化中的作用，这是一种已知与弹性蛋白降解和重塑相关的病理过程。在体外用弹性蛋白降解产物和转化生长因子 (TGF)-β 1（通常存在于恶化的基质环境中的因子）处理大鼠真皮成纤维细胞。通过逆转录酶-聚合酶链式反应、酶联免疫吸附测定、免疫荧光和钙沉积的 von Kossa 染色在基因和蛋白质水平上监测细胞变化。到第 21 天，在弹性蛋白降解产物和 TGF-β 1 单独存在的情况下，形成了多细胞钙化结节，当一起使用时，形成的程度显着更大。在矿化之前，细胞表达 α-平滑肌肌动蛋白和大量 I 型胶原蛋白和基质金属蛋白酶 2，这是肌成纤维细胞的特征，是组织重塑和修复的关键要素。受刺激的细胞表达的核心结合因子α1、骨钙素、碱性磷酸酶和骨保护素（代表性的骨调节蛋白）水平增加。对于大多数分析的蛋白质，TGF-β1 协同增强成纤维细胞对弹性蛋白降解产物的反应。总之，弹性蛋白降解产物和 TGF-β1 促进成纤维细胞中的肌成纤维细胞和成骨分化。这些结果支持弹性蛋白相关钙化涉及动态重塑事件的观点，并表明有缺陷的组织修复过程的可能性。
Our objective was to establish the role of fibroblasts in medial vascular calcification, a pathological process known to be associated with elastin degradation and remodeling. Rat dermal fibroblasts were treated in vitro with elastin degradation products and transforming growth factor (TGF)-beta 1,factors usually present in deteriorated matrix environments. Cellular changes were monitored at the gene and protein level by reverse transcriptase-polymerase chain reaction, enzyme-linked immunosorbent assay, immunofluorescence, and von Kossa staining for calcium deposits. By 21 days, multicellular calcified nodules were formed in the presence of elastin degradation products and TGF-beta 1 separately and to a significantly greater extent when used together. Before mineralization, cells expressed alpha-smooth muscle actin and large amounts of collagen type I and matrix metalloproteinase-2, characteristic features of myofibroblasts, key elements in tissue remodeling and repair. Stimulated cells expressed increased levels of core-binding factor alpha 1, osteocalcin, alkaline phosphatase, and osteoprotegerin, representative bone-regulating proteins. For most proteins analyzed, TGF-beta 1 synergistically amplified responses of fibroblasts to elastin degradation products. In conclusion, elastin degradation products and TGF-beta 1 promote myofibroblastic and osteogenic differentiation in fibroblasts. These results support the idea that elastin-related calcification involves dynamic remodeling events and suggest the possibility of a defective tissue repair process.