VE-cadherin regulates migration inhibitory factor synthesis and release
VE-cadherin regulates migration inhibitory factor synthesis and release
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VE-钙粘蛋白调节迁移抑制因子的合成和释放
DOI：
10.1007/s00011-019-01270-8
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发表时间：
2019-10-01
影响因子：
6.7
通讯作者：
Zhang, Ruyuan
中科院分区：
文献类型：
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作者：
Li, Ranran;Li, Lei;Zhang, Ruyuan
血管内皮（VE）-钙粘蛋白介导的粘附连接对维持内皮完整性至关重要。除了其嗜同性细胞间粘附的作用外，VE-钙粘蛋白还具有由外向内信号传导的作用，其对血管生理学具有功能性后果。然而，这些信号的性质仍然没有完全understood.Materials and methodsHuman umbilical vein endothelial cells（HUVECs）用于细胞培养实验。用VE-钙粘蛋白功能阻断抗体BV 9（50 μg/ml）或IgG对照处理汇合的HUVEC。抗体阵列用于筛选上清液中的细胞因子/趋化因子。对于VE-钙粘蛋白敲低，使用siRNA转染。ELISA、Western印迹和qRT-PCR用于确认筛选的细胞因子/趋化因子的表达。为了探索可能的机制，检测Scr磷酸化并使用Scr抑制剂PP 2（1 μM）。为了研究这些发现的体内相关性，将BV 9和指示的中和抗体注射到小鼠中，然后分别通过伊文思蓝测定和肺组织H&E检查肺血管渗漏和炎症。我们首先发现功能阻断抗体BV 9对VE-钙粘蛋白介导的粘附的破坏触发了迁移抑制因子（MIF）的释放。用VE-钙粘蛋白阻断抗体和siRNA技术的ELISA进一步证实了这种VE-钙粘蛋白介导的MIF释放是由于Src激酶激活介导的MIF mRNA表达增强。另外，VE-钙粘蛋白功能阻断抗体诱导的肺血管渗漏可通过中和MIF而部分减轻。结论VE-钙粘蛋白通过Src激酶调节MIF的合成和释放。我们的数据提供了额外的证据的概念，VE-钙粘蛋白转移细胞内信号，以协调状态的细胞-细胞粘附与基因表达。
ObjectiveVascular endothelial (VE)-cadherin-mediated adherens junction is critical to maintain endothelial integrity. Besides its role of homophilic intercellular adhesion, VE-cadherin also has a role of outside-in signaling with functional consequences for vascular physiology. However, the nature of these signals remains not completely understood.Materials and methodsHuman umbilical vein endothelial cells (HUVECs) were used in cell culture experiments. Confluent HUVECs were treated with VE-cadherin function-blocking antibodies BV9 (50 μg/ml) or IgG control. Antibody array was used to screen for cytokine/chemokine in supernatant. For VE-cadherin knockdown, siRNA transfection was used. ELISA, Western blot, and qRT-PCR were used to confirm the expression of screened cytokine/chemokine. To explore the possible mechanisms, Scr phosphorylation was detected and Scr inhibitor PP2 (1 μM) was used. To investigate in vivo relevance of the findings, BV9 and the indicated neutralizing antibodies were injected into mice and then lung vascular leak and inflammation were examined by Evans blue assay and lung tissue H&E, respectively.ResultsUsing a non-biased, high-throughout human cytokine/chemokine antibody array, we first found that disruption of VE-cadherin-mediated adhesion by function-blocking antibody BV9 triggered the release of migration inhibitory factor (MIF). This VE-cadherin-mediated release of MIF further confirmed by ELISA with both VE-cadherin blocking antibody and siRNA technique was due to enhanced expression of MIF mRNA, which was mediated by Src kinase activation. In addition, in vivo lung vascular leak induced by VE-cadherin function-blocking antibody was partly alleviated by neutralizing MIF.ConclusionsVE-cadherin regulates MIF synthesis and release via Src kinase. Our data provide additional evidence to the concept that VE-cadherin transfers intracellular signals to coordinate the state of cell–cell adhesion with gene expression.