Abnormal activation of calpain and protein kinase Cα promotes a constitutive release of matrix metalloproteinase 9 in peripheral blood mononuclear cells from cystic fibrosis patients
Abnormal activation of calpain and protein kinase Cα promotes a constitutive release of matrix metalloproteinase 9 in peripheral blood mononuclear cells from cystic fibrosis patients
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DOI：
10.1016/j.abb.2016.06.015
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发表时间：
2016-08-15
影响因子：
3.9
通讯作者：
Melloni, Edon
中科院分区：
文献类型：
--
作者：
Averna, Monica;Bavestrello, Margherita;Melloni, Edon
基质金属蛋白酶9（MMP 9）在生理学上参与细胞外基质成分的重塑，但在几种人类病理学中观察到其异常释放。我们在这里报告，外周血单个核细胞（PBMC），从囊性纤维化（CF）患者分离F508 del-囊性纤维化跨膜传导调节因子（CFTR）纯合子，组成型表达和释放MMP 9的高速率，由于其细胞内Ca 2+稳态的改变。这种自发和持续的MMP 9分泌可能有助于这种蛋白酶在CF患者的液体中的积累。相反，在从健康供体分离的PBMC中，MMP 9的表达和分泌是不可检测的，但可以在培养12小时后通过旁分泌刺激诱发，旁分泌刺激也促进[Ca 2 +]（i）的增加。我们还表明，在CF和对照PBMC的钙依赖性MMP 9分泌介导的钙蛋白酶和蛋白激酶C α（PKC α）的伴随激活，MMP 9的表达涉及细胞外信号调节蛋白激酶1和2（ERK 1/2）磷酸化。我们的结果得到以下事实的支持：抑制Ca 2+进入或螯合[Ca 2 +]（i）以及抑制信号通路的单个组分或恢复CFTR活性都促进MMP 9分泌的减少。(C)2016 Elsevier Inc. All rights reserved.
Matrix metalloproteinase 9 (MMP9) is physiologically involved in remodeling the extracellular matrix components but its abnormal release has been observed in several human pathologies. We here report that peripheral blood mononuclear cells (PBMCs), isolated from cystic fibrosis (CF) patients homozygous for F508del-cystic fibrosis transmembrane conductance regulator (CFTR), express constitutively and release at high rate MMP9 due to the alteration in their intracellular Ca2+ homeostasis. This spontaneous and sustained MMP9 secretion may contribute to the accumulation of this protease in fluids of CF patients. Conversely, in PBMCs isolated from healthy donors, expression and secretion of MMP9 are undetectable but can be evoked, after 12 h of culture, by paracrine stimulation which also promotes an increase in [Ca2+](i). We also demonstrate that in both CF and control PBMCs the Ca2+-dependent MMP9 secretion is mediated by the concomitant activation of calpain and protein kinase C alpha (PKC alpha), and that MMP9 expression involves extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) phosphorylation. Our results are supported by the fact that either the inhibition of Ca2+ entry or chelation of [Ca2+](i) as well as the inhibition of single components of the signaling pathway or the restoration of CFTR activity all promote the reduction of MMP9 secretion. (C) 2016 Elsevier Inc. All rights reserved.