Defining the involvement of proteinases in pemphigus vulgaris: Evidence of matrix metalloproteinase-9 overexpression in experimental models of disease
Defining the involvement of proteinases in pemphigus vulgaris: Evidence of matrix metalloproteinase-9 overexpression in experimental models of disease
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DOI：
10.1002/jcp.20997
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发表时间：
2007-07-01
影响因子：
5.6
通讯作者：
Lanza, Alessandro
中科院分区：
文献类型：
--
作者：
Cirillo, Nicola;Lanza, Michele;Lanza, Alessandro
寻常天疱疮（PV）棘层松解是一个复杂的现象，其中许多因素的合作。已知PV血清调节重要的细胞事件，包括激酶活性、转录调节和蛋白酶表达。事实上，由PV血清触发的细胞信号转导可能诱导蛋白酶上调，可能导致表皮粘附破坏，并最终导致水疱形成。在这里，我们试图通过使用体内和体外PV模型来研究这一假设。对小鼠皮肤组织的微阵列分析表明，细胞外蛋白酶及其抑制剂之间的平衡朝着增强的蛋白水解活性在PV新生小鼠模型，至少在转录水平上。相反，编码细胞粘附蛋白的基因显著下调。然后在体外角质形成细胞单层和皮肤器官培养物中研究PV血清对蛋白水平的影响，重点是基质金属蛋白酶（MMP）9的表达和活性。通过Western印迹，酶谱，和活细胞免疫荧光研究，我们表明，MMP-9早期过度表达的角质形成细胞暴露于PV血清，随后分泌在培养基中。然而，我们未能证明MMP-9的细胞外活化，因为在无血清培养上清液中发现其92 kDa的无活性形式。综上所述，我们的数据表明，蛋白酶的表达，特别是MMP-9，是由PV血清调制，并与IPV棘层松解。
Pemphigus vulgaris (PV) acantholysis represents a complex phenomenon wherein a number of factors cooperates. PV serum is known to modulate important cellular events, including kinase activity, transcriptional regulation, and proteinase expression. Indeed, transduction of signals to the cell triggered by PV serum may induce proteinase up-regulation potentially responsible for disruption of epidermal adhesion and, ultimately, blister formation. Here, we sought to investigate this hypothesis by using both in vivo and in vitro models of PV. Microarray analysis on mouse skin tissues suggested that the equilibrium between extracellular proteinases and their inhibitors moved towards enhanced proteolytic activity in PV neonatal mouse model, at least on the transcriptional level. Conversely, genes codifying cell adhesion proteins were dramatically down-regulated. The effects of PV serum on the protein level were then studied in vitro both in keratinocyte monolayers and skin organ cultures focusing on matrix metalloproteinase (MMP) 9 expression and activity. By means of Western blotting, zymography, and living cell immunofluorescence studies, we showed that MMP-9 was early overexpressed in keratinocytes exposed to PV serum, and subsequently secreted in the culture medium. However, we failed to demonstrate extracellular activation of MMP-9, since it was found in its 92 kDa inactive form in serum-free culture supernatants. Taken together, our data demonstrated that proteinase expression, particularly of MMP-9, is modulated by PV serum and associated with IPV acantholysis.