Role of Src in Vascular Hyperpermeability Induced by Advanced Glycation End Products.
Role of Src in Vascular Hyperpermeability Induced by Advanced Glycation End Products.
复制标题
Src 在高级糖基化终产物诱导的血管通透性过高中的作用
DOI：
10.1038/srep14090
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发表时间：
2015-09-18
影响因子：
4.6
通讯作者：
Guo X
中科院分区：
文献类型：
--
作者：
Zhang W;Xu Q;Wu J;Zhou X;Weng J;Xu J;Wang W;Huang Q;Guo X
晚期糖基化终产物(AGEs)刺激引起的微血管屏障的破坏导致了糖尿病相关的血管病变。为了研究Src在AGE诱导的血管高通透性中的作用及其与Moesin、VE-钙粘附素和粘着斑激酶(FAK)的关系，我们证实了AGE诱导了Src的磷酸化，导致HUVECs通透性增加。AGE处理后，过表达的细胞显示出更高的通透性，并伴有更明显的F-肌动蛋白重排。仅用pcDNA3/FLAG-SrcY530F激活Src就复制了这些效应。用siRNA、PP2或pcDNA3/FLAG-SrcK298M抑制Src可消除上述作用。从AGEs受体(RAGE)基因敲除小鼠分离的肺微血管内皮细胞(PMVECs)可降低AGE基因敲除小鼠肺组织中Src的磷酸化水平，减轻AGE治疗后的屏障功能障碍。体内研究表明，AGE可使小鼠肠系膜小静脉分泌的右旋糖酐增多。在RAGE基因敲除或PP2预处理的小鼠中，这种作用被减弱。上调Src活性可诱导Moesin的磷酸化以及VE-cadherin的激活和解离，而下调Src则取消这些作用。在AGE刺激的人脐静脉内皮细胞中，FAK也被证明与Src相互作用。我们的研究表明，通过分别磷酸化Moesin、VE-cadherin和FAK，Src在AGE诱导的微血管高通透性中起关键作用。
The disruption of microvascular barrier in response to advanced glycation end products (AGEs) stimulation contributes to vasculopathy associated with diabetes mellitus. Here, to study the role of Src and its association with moesin, VE-cadherin and focal adhesion kinase (FAK) in AGE-induced vascular hyperpermeability, we verified that AGE induced phosphorylation of Src, causing increased permeability in HUVECs. Cells over-expressed Src displayed a higher permeability after AGE treatment, accompanied with more obvious F-actin rearrangement. Activation of Src with pcDNA3/flag-SrcY530F alone duplicated these effects. Inhibition of Src with siRNA, PP2 or pcDNA3/flag-SrcK298M abolished these effects. The pulmonary microvascular endothelial cells (PMVECs) isolated from receptor for AGEs (RAGE)-knockout mice decreased the phosphorylation of Src and attenuated the barrier dysfunction after AGE-treatment. In vivo study showed that the exudation of dextran from mesenteric venules was increased in AGE-treated mouse. This was attenuated in RAGE knockout or PP2-pretreated mice. Up-regulation of Src activity induced the phosphorylation of moesin, as well as activation and dissociation of VE-cadherin, while down-regulation of Src abolished these effects. FAK was also proved to interact with Src in HUVECs stimulated with AGEs. Our studies demonstrated that Src plays a critical role in AGE-induced microvascular hyperpermeability by phosphorylating moesin, VE-cadherin, and FAK respectively.