Ischemia-induced cleavage of cadherins in NRK cells requires MT1-MMP (MMP-14)
Ischemia-induced cleavage of cadherins in NRK cells requires MT1-MMP (MMP-14)
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DOI：
10.1152/ajprenal.00179.2005
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发表时间：
2006-01-01
影响因子：
4.2
通讯作者：
Parrish, AR
中科院分区：
文献类型：
--
作者：
Covington, MD;Burghardt, RC;Parrish, AR
缺血诱导NRK细胞中钙粘附素的切割需要MT1-MMP14(MMP14)。Am J Physiol Renal Physiol 290：F43-F51,2006.2005年8月2日首次出版；DOI：10.1152/ajprenal.00179.2005。-缺血是急性肾功能衰竭的主要原因，急性肾功能衰竭是一种与高发病率和死亡率相关的疾病。近端小管细胞间黏附的破坏与ARF有关，尽管其分子机制尚不清楚(S)。我们以前的研究表明，缺血与NRK细胞中钙粘附素的切割和丢失有关，可能是由于基质金属蛋白酶(MMP)(7)所致。在目前的研究中，确定了E-钙粘蛋白切割和N-钙粘蛋白丢失所需的基质金属蛋白酶。针对一些可溶性MMPs的化学抑制剂(1、2、3、8、9)不能完全减弱缺血诱导的钙粘附素丢失。在缺血条件下，活性膜型(MT)1-MMPs表达增加，而MMP2蛋白表达减少。纤维连接蛋白上的电镀细胞可保护缺血诱导的钙粘附素丢失，有趣的是，在纤维连接蛋白涂层的培养皿中，缺血细胞中活跃的MT1-基质金属蛋白酶水平没有增加。此外，L细胞稳定表达E-(LE)或N-钙粘蛋白(LN)，但缺乏MT1-MMPs的表达，对缺血诱导的钙粘附素丢失具有抵抗作用。MT1-MMPs在缺血诱导的钙粘附素丢失中的作用可以通过中和抗体阻断MT1-MMPs的活性或用保护全长E-和N-Cadherin的shRNA构建物来证实。使用shRNA构建物抑制MT1-MMPs的表达，可以消除缺血引起的钙粘蛋白功能中断，并保留细胞与细胞之间的接触。这些结果表明，缺血诱导活化的MT1-MMPs表达增加，继而破坏钙粘附素/连环蛋白复合体，提示MT1-MMPs在缺血诱导的ARF中起一定作用。
Ischemia-induced cleavage of cadherins in NRK cells requires MT1-MMP (MMP-14). Am J Physiol Renal Physiol 290: F43-F51, 2006. First published August 2, 2005; doi:10.1152/ajprenal.00179.2005.- Ischemia is a leading cause of acute renal failure (ARF), a disease associated with high morbidity and mortality. Disruption of intercellular adhesion in the proximal tubules is linked to ARF, although the molecular mechanism(s) remains unclear. Our previous studies showed that ischemia is associated with cadherin cleavage and loss in NRK cells, putatively due to a matrix metalloproteinase (MMP) (7). In the current studies, a MMP required for E-cadherin cleavage and N-cadherin loss was identified. Chemical inhibitors against a number of soluble MMPs ( 1, 2, 3, 8, 9) failed to completely attenuate ischemia-induced cadherin loss. Under ischemic conditions, there was an increase in active membrane-type (MT) 1-MMP but a decrease in MMP-2 protein expression. Plating cells on fibronectin protected against ischemia-induced loss of cadherins and, interestingly, no increase in active MT1-MMP levels was seen in ischemic cells on fibronectin-coated dishes. In addition, L cells stably expressing E- (LE) or N- cadherin (LN), but lacking MT1-MMP expression, were resistant to ischemia-induced cadherin loss. The role of MT1-MMP in ischemia-induced cadherin loss was confirmed by either blocking MT1-MMP activity with a neutralizing antibody or expression with shRNA constructs which protected full-length E- and N- cadherin during ischemia. Using shRNA constructs to suppress MT1-MMP expression, ischemia-induced disruption of cadherin function was ablated, and cell-cell contacts were preserved. These results demonstrate that ischemia induces increased expression of active MT1-MMP and subsequent disruption of cadherin/catenin complexes, implying that MT1-MMP plays a role in ischemia-induced ARF.