Elastin calcification in the rat subdermal model is accompanied by up-regulation of degradative and osteogenic cellular responses
Elastin calcification in the rat subdermal model is accompanied by up-regulation of degradative and osteogenic cellular responses
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DOI：
10.2353/ajpath.2006.050338
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发表时间：
2006-02-01
影响因子：
6
通讯作者：
Vyavahare, NR
中科院分区：
文献类型：
--
作者：
Lee, JS;Basalyga, DM;Vyavahare, NR
血管弹性蛋白钙化发生在动脉硬化、肾功能衰竭、糖尿病和血管移植植入患者中。我们假设病理性弹性蛋白钙化与退行性和成骨机制有关。为了验证这一假设，通过定量实时逆转录-聚合酶链反应和特异性蛋白检测，在大鼠皮下钙化模型中检测了与弹性蛋白降解和成骨相关的基因和蛋白质的时间表达。幼年大鼠皮下植入纯化弹性蛋白后，随着成纤维细胞和巨噬细胞的浸润，呈时间依赖性的进行性钙化。逆转录-聚合酶链反应分析显示基质金属蛋白酶（MMP-2、MMP-9）和转化生长因子- β 1基因的相对表达水平与钙化同步升高。明胶酶谱分析显示，在早期时间点MMP活性较强，这与高水平的可溶性弹性蛋白肽有关。核心结合因子α -1（一种成骨细胞特异性转录因子）的基因表达与弹性蛋白钙化同时增加，与植入后3天相比，21天的表达量增加了9.5倍。同样，骨标志物骨桥蛋白和碱性磷酸酶的mRNA水平也逐渐升高，但骨钙素水平保持不变。我们得出结论，退行性和成骨过程可能参与弹性蛋白钙化。
Calcification of vascular elastin occurs in patients with arteriosclerosis, renal failure, diabetes, and vascular graft implants. We hypothesized that pathological elastin calcification is related to degenerative and osteogenic mechanisms. To test this hypothesis, the temporal expression of genes and proteins associated with elastin degradation and osteogenesis was examined in the rat subdermal calcification model by quantitative real-time reverse transcription-polymerase chain reaction and specific protein assays. Purified elastin implanted subdermally in juvenile rats exhibited progressive calcification in a time-dependent manner along with fibroblast and macrophage infiltration. Reverse transcription-polymerase chain reaction analysis showed that relative gene expression levels of matrix metalloproteinases (MMP-2 and MMP-9) and transforming growth factor-beta 1 were increased in parallel with calcification. Gelatin zymography showed strong MMP activities at early time points, which were associated with high levels of soluble elastin peptides. Gene expression of core binding factor alpha-1, an osteoblast-specific transcription factor, increased in parallel with elastin calcification and attained similar to 9.5-fold higher expression at 21 days compared to 3 days after implantation. Similarly, mRNA levels of the bone markers osteopontin and alkaline phosphatase also increased progressively, but osteocalcin levels remained unchanged. We conclude that degenerative and osteogenic processes may be involved in elastin calcification.