Fibroblasts from patients affected by Pseudoxanthoma elasticum exhibit an altered PPi metabolism and are more responsive to pro-calcifying stimuli
Fibroblasts from patients affected by Pseudoxanthoma elasticum exhibit an altered PPi metabolism and are more responsive to pro-calcifying stimuli
复制标题
DOI：
10.1016/j.jdermsci.2013.12.008
复制
发表时间：
2014-04-01
影响因子：
4.6
通讯作者：
Quaglino, Daniela
中科院分区：
文献类型：
--
作者：
Boraldi, Federica;Annovi, Giulia;Quaglino, Daniela
背景：弹性纤维性假黄瘤是一种以软结缔组织进行性钙化为特征的遗传性疾病。其发病机制至今仍难以确定。在PXE真皮成纤维细胞中，除了维生素K依赖性抑制剂基质Gla蛋白（MGP）的羧化受损外，我们还证明了碱性磷酸酶活性的上调。根据这些数据，我们认为钙和磷酸盐代谢可能发生局部改变，这两种途径在基质钙化的发生中起协同作用。目的：本研究旨在更好地探索培养的PXE成纤维细胞与对照细胞相比，是否表现出修饰的无机焦磷酸盐（PPi）代谢，并且对促钙化刺激更敏感。将从健康个体和PXE患者分离的原代人真皮成纤维细胞在标准培养基和促钙化培养基中培养不同时间点。分别采用qRT-PCR和Western blot检测ANKH/ANKH、ENPP 1/PC 1、ALPL/TNAP、SPP 1/OPN的表达。TNAP活性测定分光光度分析，而钙化进行了研究，通过光学和电子显微镜，以及通过microanalytical techniques.Results：在存在的促钙化刺激，真皮成纤维细胞改变其表型有利于基质矿化。特别地，ENPP 1/PC 1和SPP 1/OPN表达以及TNAP活性在对照和PXE成纤维细胞中表达不同。此外，在病理细胞之间的比例有利于和减少PPi的可用性的因素表现出更明显的转变对一个前钙化balancy.Conclusion：PXE成纤维细胞更容易受到前钙化的刺激，并在这些细胞的PPi代谢的改变有助于基质钙化。(c)2014年日本皮肤病研究学会。由Elsevier爱尔兰有限公司出版。保留所有权利。
Background: Pseudoxanthoma elasticum (PXE) is a genetic disorder characterized by progressive calcification of soft connective tissues. The pathogenesis is still hard to pin down. In PXE dermal fibroblasts, in addition to impaired carboxylation of the vitamin K-dependent inhibitor matrix Gla protein (MGP), we have also demonstrated an up-regulation of alkaline phosphatase activity. In the light of these data we have suggested that both calcium and phosphate metabolism might be locally altered, both pathways acting in synergy on the occurrence of matrix calcification.Objective: This study aims to better explore if cultured PXE fibroblasts, compared to control cells, exhibit a modified inorganic pyrophosphate (PPi) metabolism and are more responsive to pro-calcifying stimuli.Methods: Primary human dermal fibroblasts isolated from healthy individuals and from PXE patients were cultured for different time points in standard and in pro-calcifying media. The expression of ANKH/ANKH, ENPP1/PC1, ALPL/TNAP, SPP1/OPN was evaluated by qRT-PCR and Western blot, respectively. TNAP activity was measured by spectrophotometric analyses, whereas calcification was investigated by light and electron microscopy as well as by micro-analytical techniques.Results: In the presence of pro-calcifying stimuli, dermal fibroblasts alter their phenotype favouring matrix mineralization. In particular, ENPP1/PC1 and SPP1/OPN expression, as well as TNAP activity, was differently expressed in control and in PXE fibroblasts. Moreover, in pathologic cells the ratio between factors favouring and reducing PPi availability exhibits a more pronounced shift towards a pro-calcifying balance.Conclusion: PXE fibroblasts are more susceptible to pro-calcifying stimuli and in these cells an altered PPi metabolism contributes to matrix calcification. (c) 2014 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.