High-mobility group box 1 impairs airway epithelial barrier function through the activation of the RAGE/ERK pathway.
High-mobility group box 1 impairs airway epithelial barrier function through the activation of the RAGE/ERK pathway.
复制标题
高迁移率组盒 1 通过激活 RAGE/ERK 通路损害气道上皮屏障功能
DOI：
10.3892/ijmm.2016.2537
复制
发表时间：
2016-05
影响因子：
5.4
通讯作者：
Cai S
中科院分区：
文献类型：
--
作者：
Huang W;Zhao H;Dong H;Wu Y;Yao L;Zou F;Cai S
最近的研究表明，高迁移率族蛋白1(HMGB1)和晚期糖基化终产物受体(RAGE)参与了哮喘的发病。然而，HMGB1/RAGE轴的激活是否介导了呼吸道上皮屏障功能障碍尚不清楚。因此，本研究的目的是研究HMGB1及其与白介素1β的协同作用对呼吸道上皮屏障特性的影响。我们通过培养气液界面16HBE细胞，模拟分化的上皮细胞，评价了重组人HMGB1单独或联合IL-1β对离子和大分子屏障通透性的影响。Western印迹分析和免疫荧光染色检测主要连接蛋白E-钙粘素、β-连环蛋白、封闭蛋白和Claudin-1的水平和结构。此外，我们检测了RAGE中和抗体和丝裂原活化蛋白激酶(MAPK)抑制剂对上皮屏障特性的影响，以阐明相关的机制。HMGB1以剂量和时间依赖的方式增加FITC-葡聚糖通透性，但抑制上皮抵抗。HMGB1介导的屏障高通透性伴随着细胞-细胞接触的破坏、occludin和claudin-1的选择性下调以及E-cad和β-catenin的重新分布。HMGB1与IL-1β协同作用可诱导类似但更强的屏障高通透性，并诱导连接蛋白的破坏。此外，HMGB1激活RAGE/细胞外信号相关激酶(ERK)1/2信号通路，与16HBE细胞屏障功能障碍相关。抗RAGE抗体和ERK1/2抑制剂U0126可减轻HMGB1介导的屏障通透性改变，恢复occludin和claudin-1的表达水平，并阻止E-钙粘素和β-catenin的重新分布。综上所述，我们的研究结果表明，HMGB1能够诱导对上皮屏障功能的强烈影响，并且RAGE/ERK1/2是参与连接蛋白形成和上皮屏障功能障碍之间的串扰的关键信号通路。
Recent studies have indicated that high-mobility group box 1 protein (HMGB1) and the receptor for advanced glycation end-products (RAGE) contribute to the pathogenesis of asthma. However, whether the activation of the HMGB1/RAGE axis mediates airway epithelial barrier dysfunction remains unknown. Thus, the aim of this study was to examine the effects of HMGB1 and its synergistic action with interleukin (IL)-1β on airway epithelial barrier properties. We evaluated the effects of recombinant human HMGB1 alone or in combination with IL-1β on ionic and macromolecular barrier permeability, by culturing air-liquid interface 16HBE cells with HMGB1 to mimic the differentiated epithelium. Western blot analysis and immunofluorescence staining were utilized to examine the level and structure of major junction proteins, namely E-cadherin, β-catenin, occludin and claudin-1. Furthermore, we examined the effects of RAGE neutralizing antibodies and mitogen-activated protein kinase (MAPK) inhibitors on epithelial barrier properties in order to elucidate the mechanisms involved. HMGB1 increased FITC-dextran permeability, but suppressed epithelial resistance in a dose-and time-dependent manner. HMGB1-mediated barrier hyperpermeability was accompanied by a disruption of cell-cell contacts, the selective downregulation of occludin and claudin-1, and the redistribution of E-cadherin and β-catenin. HMGB1 in synergy with IL-1β induced a similar, but greater barrier hyperpermeability and induced the disruption of junction proteins. Furthermore, HMGB1 elicited the activation of the RAGE/extracellular signal-related kinase (ERK)1/2 signaling pathway, which correlated with barrier dysfunction in the 16HBE cells. Anti-RAGE antibody and the ERK1/2 inhibitor, U0126, attenuated the HMGB1-mediated changes in barrier permeability, restored the expression levels of occludin and claudin-1 and pevented the redistribution of E-cadherin and β-catenin. Taken together, the findings of our study demonstrate that HMGB1 is capable of inducing potent effects on epithelial barrier function and that RAGE/ERK1/2 is a key signaling pathway involved in the crosstalk between formations of junction proteins and epithelial barrier dysfunction.