南方医科大学学报2017，Vol.37Issue(6)：737-743,7.DOI:10.3969/j.issn.1673-4254.2017.06.04
表皮生长因子受体参与屋尘螨诱导的气道上皮屏障破坏的机制
Role of epidermal growth factor receptor in house dust mite-induced airway epithelial barrier dysfunction
摘要
Abstract
Objective To investigate the role of epidermal growth factor receptor (EGFR) signaling pathway in bronchial epithelial actin stress fiber (F-actin) rearrangement induced by house dust mite (HDM). Methods Normal human bronchial epithelial cells (16HBE) were stimulated with HDM with or without pretreatment with AG-1478, an EGFR inhibitor. The levels of phospho(p)-EGFR, F-actin, E-cadherin and β-catenin in the cell cultures were detected with Western blotting. The localizations of F-actin, E-cadherin and β-catenin in the bronchial epithelial cells were determined with immunofluorescence assay, and the transmembrane electrical resistance (TER) and FITC-dextran flux (FITC-DX) in the cells were measured to assess the barrier function of the bronchial epithelia. Results HDM stimulation of the cells for 10 min resulted in significantly increased p-EGFR expression (P<0.05) without causing obvious changes in the expression of E-cadherin (P>0.05) orβ-catenin (P>0.05). Immunofluorescence assay revealed delocalization of E-cadherin and β-catenin in HDM-treated 16HBE cells, shown by their diffusion from the cell membrane to the cytoplasm. In HDM-treated cells, the TER was significantly decreased to (70.00±4.33)%and the FITC-DX was significantly increased to (115.98±4.34)%;Inhibition of EGFR reversed the delocalization of E-cadherin and β-catenin, improved the TER to (90.00 ± 3.75)% and lowered the FITC-DX to (101.10 ± 2.10)%. HDM induced increased expression and rearrangement of F-actin, which was obviously inhibited by pretreatment of the cells with AG-1478 (P<0.05). Conclusion EGFR signaling pathway mediates HDM-induced F-actin rearrangement in human bronchial epithelial cells to contribute to epithelial barrier dysfunction.关键词
表皮生长因子受体/肌动蛋白应力纤维/屋尘螨/气道上皮/屏障破坏Key words
epidermal growth factor receptor/F-actin/house dust mite/airway epithelia/barrier dysfunction引用本文复制引用
乐艳青,董航明,王燕红,赵海金,蔡绍曦..表皮生长因子受体参与屋尘螨诱导的气道上皮屏障破坏的机制[J].南方医科大学学报,2017,37(6):737-743,7.基金项目
国家自然科学基金(81670026,81500023,81470228, 81270087,81270089) （81670026,81500023,81470228, 81270087,81270089）
国家重点基础研究发展计划"973"计划(2012CB518203) （2012CB518203）
2016 年" 精 准 医 学 研 究 "重 点 专 项(2016YFC0905800) （2016YFC0905800）
广东省科技计划项目(2015A030313236, 2014A030310325) Supported by National Natural Science Foundation of China (81670026, 81500023,81470228,81270087,81270089). （2015A030313236, 2014A030310325）
