Drug Discovery Research Center, Southwest Medical University, 319 Zhongshan Road, Luzhou 646000, China.
First Clinical College of Medicine, Southern Medical University, Guangzhou 510515, China.
Sensors (Basel). 2017 Mar 30;17(4):722. doi: 10.3390/s17040722.
Endothelial functional dysregulation and barrier disruption contribute to the initiation and development of sepsis. The receptor for advanced glycation end products (RAGE) has been demonstrated to be involved in the pathogenesis of sepsis. The present study aimed to investigate the role of RAGE in lipopolysaccharide (LPS)-induced nuclear factor-κB (NF-κB) activation in endothelial cells and the consequent endothelial hyperpermeability. LPS-induced upregulation of RAGE protein expression in human umbilical vein endothelial cells (HUVECs) was detected by western blotting. Activation of NF-κB was revealed using western blotting and immunofluorescent staining. LPS-elicited endothelial hyperpermeability was explored by transendothelial electrical resistance (TER) assay and endothelial monolayer permeability assay. The blocking antibody specific to RAGE was used to confirm the role of RAGE in LPS-mediated NF-κB activation and endothelial barrier disruption. We found that LPS upregulated the protein expression of RAGE in a dose- and time-dependent manner in HUVECs. Moreover, LPS triggered a significant phosphorylation and degradation of IκBα, as well as NF-κB p65 nuclear translocation. Moreover, we observed a significant increase in endothelial permeability after LPS treatment. However, the RAGE blocking antibody attenuated LPS-evoked NF-κB activation and endothelial hyperpermeability. Our results suggest that RAGE plays an important role in LPS-induced NF-κB activation and endothelial barrier dysfunction.
内皮功能失调和屏障破坏导致脓毒症的发生和发展。已证明晚期糖基化终产物（RAGE）受体参与脓毒症的发病机制。本研究旨在探讨 RAGE 在脂多糖（LPS）诱导的内皮细胞核因子-κB（NF-κB）激活及其随后的内皮通透性增加中的作用。通过 Western blot 检测 LPS 诱导的人脐静脉内皮细胞（HUVEC）中 RAGE 蛋白表达的上调。通过 Western blot 和免疫荧光染色揭示 NF-κB 的激活。通过跨内皮电阻（TER）测定和内皮单层通透性测定探讨 LPS 诱导的内皮通透性增加。使用针对 RAGE 的阻断抗体来确认 RAGE 在 LPS 介导的 NF-κB 激活和内皮屏障破坏中的作用。我们发现 LPS 以剂量和时间依赖的方式在上皮细胞中上调 RAGE 蛋白表达。此外，LPS 触发了 IκBα的显著磷酸化和降解，以及 NF-κB p65 的核转位。此外，我们观察到 LPS 处理后内皮通透性显著增加。然而，RAGE 阻断抗体减弱了 LPS 诱导的 NF-κB 激活和内皮通透性增加。我们的结果表明，RAGE 在 LPS 诱导的 NF-κB 激活和内皮屏障功能障碍中发挥重要作用。
