Modulatory role of focal adhesion kinase in regulating human pulmonary arterial endothelial barrier function
Modulatory role of focal adhesion kinase in regulating human pulmonary arterial endothelial barrier function
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DOI：
10.1113/jphysiol.2001.013289
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发表时间：
2002-03-15
影响因子：
5.5
通讯作者：
Malik, AB
中科院分区：
文献类型：
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作者：
Mehta, D;Tiruppathi, C;Malik, AB
整合素受体与细胞外基质（ECM）在粘着斑复合物处相互作用产生的粘附力可调节内皮细胞形状，从而调节内皮屏障功能。我们使用培养的人肺动脉内皮细胞研究了由整合素信号激活的粘着斑激酶（FAK）在调节细胞形状中的作用。我们使用FAK反义寡核苷酸（靶向FAK mRNA的3 '非翻译区（5'-CTCTGGTTGATGGGATTG-3 '））来确定FAK在凝血酶诱导的内皮通透性增加机制中的作用。反义FAK表达的减少增强了凝血酶诱导的跨内皮电阻的降低（模拟转染细胞降低-43 +/-1%，正义转染细胞降低-40 +/-4%，而反义转染细胞降低-60 +/-3%）。FAK表达的减少也延长了电阻的下降，并阻止了在对照内皮细胞中观察到的恢复。因此，凝血酶诱导的渗透性增加在FAK表达不存在的情况下既更大又减弱。用latrunculin-A抑制肌动蛋白聚合可防止FAK移位到粘着斑和FAK及桩蛋白的酪氨酸磷酸化，并同时使凝血酶诱导的电阻降低约50%。因此，FAK对内皮屏障功能的调节作用依赖于肌动蛋白聚合。在肌动蛋白缆索的引导下，FAK移位至内皮细胞中的粘着斑复合物，随后FAK相关蛋白的活化用于逆转由炎症介质如凝血酶引起的内皮屏障功能的降低。
The adhesive force generated by the interaction of integrin receptors with extracellular matrix (ECM) at the focal adhesion complex may regulate endothelial cell shape, and thereby the endothelial barrier function. We studied the role of focal adhesion kinase (FAK) activated by integrin signalling in regulating cell shape using cultured human pulmonary artery endothelial cells. We used FAK antisense oligonucleotide (targeted to the 3'-untranslated region of FAK mRNA (5'-CTCTGGTTGATGGGATTG-3') to determine the role of FAK in the mechanism of thrombin-induced increase in endothelial permeability. Reduction in FAK expression by the antisense augmented the thrombin-induced decrease in transendothelial electrical resistance (decrease in mock transfected cells of -43 +/- 1% and in sense-transfected cells of -40 +/- 4 %, compared to the decrease in antisense-transfected cells of -60 +/- 3%). Reduction in FAK expression also prolonged the drop in electrical resistance and prevented the recovery seen in control endothelial cells. Thus, the thrombin-induced increase in permeability is both greater and attenuated in the absence of FAK expression. Inhibition of actin polymerization with latrunculin-A prevented the translocation of FAK to focal adhesion sites and tyrosine phosphorylation of FAK and paxillin, and concomitantly reduced the thrombin-induced decrease in electrical resistance by similar to50%. Thus, the modulatory role of FAK on endothelial barrier function is dependent on actin polymerization. FAK translocation to focal adhesion complex in endothelial cells guided by actin cables and the consequent activation of FAK-associated proteins serve to reverse the decrease in endothelial barrier function caused by inflammatory mediators such as thrombin.