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首页> 外文期刊>Journal of Cellular Physiology >Fibrinogen induces alterations of endothelial cell tight junction proteins.
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Fibrinogen induces alterations of endothelial cell tight junction proteins.

机译:纤维蛋白原诱发内皮细胞的变化紧密连接蛋白。

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We previously showed that an elevated content of fibrinogen (Fg) increased formation of filamentous actin and enhanced endothelial layer permeability. In the present work we tested the hypothesis that Fg binding to endothelial cells (ECs) alters expression of actin-associated endothelial tight junction proteins (TJP). Rat cardiac microvascular ECs were grown in gold plated chambers of an electrical cell-substrate impedance system, 8-well chambered, or in 12-well plates. Confluent ECs were treated with Fg (2 or 4 mg/ml), Fg (4 mg/ml) with mitogen-activated protein kinase (MEK) kinase inhibitors (PD98059 or U0126), Fg (4 mg/ml) with anti-ICAM-1 antibody or BQ788 (endothelin type B receptor blocker), endothelin-1, endothelin-1 with BQ788, or medium alone for 24 h. Fg induced a dose-dependent decrease in EC junction integrity as determined by transendothelial electrical resistance (TEER). Western blot analysis and RT-PCR data showed that the higher dose of Fg decreased the contents of TJPs, occludin, zona occluden-1 (ZO-1), and zona occluden-2 (ZO-2) in ECs. Fg-induced decreases in contents of the TJPs were blocked by PD98059, U0126, or anti-ICAM-1 antibody. While BQ788 inhibited endothelin-1-induced decrease in TEER, it did not affect Fg-induced decrease in TEER. These data suggest that Fg increases EC layer permeability via the MEK kinase signaling pathway by affecting occludin, ZO-1, and ZO-2, TJPs, which are bound to actin filaments. Therefore, increased binding of Fg to its major EC receptor, ICAM-1, during cardiovascular diseases may increase microvascular permeability by altering the content and possibly subcellular localization of endothelial TJPs.
机译:我们以前显示的含量升高纤维蛋白原(Fg)增加的形成丝状肌动蛋白和增强内皮细胞层渗透率。假设Fg绑定到内皮细胞(ECs)改变actin-associated的表情内皮细胞紧密连接蛋白(TJP)。心脏微血管ECs是生长在黄金镀室的电气的细胞基质系统阻抗,8-well有房间的,或者在12-well盘子。4毫克/毫升),成品与增殖作用(4毫克/毫升)蛋白激酶激酶抑制剂(PD98059 (MEK)或U0126),成品与anti-ICAM-1抗体(4毫克/毫升)或BQ788(内皮素B型受体阻断剂),endothelin-1, endothelin-1 BQ788或媒介仅24 h。Fg诱导剂量依赖性减少电子商务连接完整性所确定的由transendothelial电阻(te)。免疫印迹分析和rt - pcr数据显示成品的高剂量减少的内容TJPs occludin、occluden-1 (ZO-1)区和区ECs occluden-2 (ZO-2)。内容TJPs PD98059被封锁,U0126或anti-ICAM-1抗体。抑制endothelin-1-induced减少三通,它并不影响Fg-induced te下降。这些数据表明,Fg增加电子商务层渗透率通过MEK激酶信号通路通过影响occludin ZO-1, ZO-2 TJPs,绑定到肌动蛋白丝。增加成品的约束力主要EC受体,ICAM-1,在心血管疾病的可能增加微血管通透性的改变内容和可能的亚细胞定位内皮TJPs。

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