p38 MAP kinase-dependent regulation of endothelial cell permeability
p38 MAP kinase-dependent regulation of endothelial cell permeability
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DOI：
10.1152/ajplung.00372.2003
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发表时间：
2004-11-01
影响因子：
4.9
通讯作者：
Verin, AD
中科院分区：
文献类型：
--
作者：
Borbiev, T;Birukova, A;Verin, AD
我们之前已经证明凝血酶通过细胞骨架激活和收缩诱导内皮细胞屏障功能障碍，并确定了内皮细胞肌球蛋白轻链激酶（MLCK）在这一过程中的重要作用。在本研究中，我们探索了p38 MAP激酶作为凝血酶介导的内皮细胞收缩反应和通透性的潜在重要酶。凝血酶以时间依赖性的方式诱导p38 MAP激酶激活，在30分钟时效果最大，这与肌动蛋白和肌球蛋白结合蛋白caldesmon磷酸化增加有关。SB-203580和显性阴性p38腺病毒载体均能显著减弱凝血酶诱导的跨内皮电阻下降。与这些数据一致，SB-203580减少了内皮中凝血酶产生的肌动蛋白应激纤维的形成。此外，显性阴性p38对凝血酶诱导的肌球蛋白轻链二磷酸化没有影响。SB-203580预处理可减弱凝血酶诱导的总磷酸化和位点特异性caldesmon磷酸化（Ser(789)）以及caldesmon-myosin复合物的解离。这些结果表明p38 MAP激酶活性和caldesmon磷酸化参与了mlck不依赖于凝血素诱导的内皮细胞通透性的调节。
We have previously shown that thrombin induces endothelial cell barrier dysfunction via cytoskeleton activation and contraction and have determined the important role of endothelial cell myosin light chain kinase (MLCK) in this process. In the present study we explored p38 MAP kinase as a potentially important enzyme in thrombin-mediated endothelial cell contractile response and permeability. Thrombin induces significant p38 MAP kinase activation in a time-dependent manner with maximal effect at 30 min, which correlates with increased phosphorylation of actin- and myosin-binding protein, caldesmon. Both SB-203580 and dominant negative p38 adenoviral vector significantly attenuated thrombin-induced declines in transendothelial electrical resistance. Consistent with these data SB-203580 decreased actin stress fiber formation produced by thrombin in endothelium. In addition, dominant negative p38 had no effect on thrombin-induced myosin light chain diphosphorylation. Thrombin-induced total and site-specific caldesmon phosphorylation (Ser(789)) as well as dissociation of caldesmon-myosin complex were attenuated by SB-203580 pretreatment. These results suggest the involvement of p38 MAP kinase activities and caldesmon phosphorylation in the MLCK-independent regulation of thrombin-induced endothelial cell permeability.