Page 38 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第42卷第6期
·790 · Journal of Nanjing Medical University（Natural Sciences） 2022年6月

·基础研究·

蛋白酶激活受体2经ROS信号调控支气管上皮间充质表型转化

王红玉，顾 浩，葛 爱，曾晓宁 ，姚 欣 *
*
南京医科大学第一附属医院呼吸与危重症医学科，江苏 南京 210029

［摘 要］ 目的：探讨蛋白酶激活受体2（protease⁃activated receptor 2，PAR2）在人支气管上皮细胞间充质转化（epithelial⁃mesen⁃
chymal transition，EMT）中的作用及可能机制。方法：16HBE 细胞予以不同浓度类胰蛋白酶（PAR2 激动剂）处理，采用划痕实
验、Transwell小室、蛋白质免疫印迹等方法观察类胰蛋白酶对细胞EMT过程的影响；通过DCFH⁃DA探针测定细胞活性氧（re⁃
active oxygen species，ROS）的水平；外源性加入ROS清除剂N⁃乙酰半胱氨酸（N⁃acety⁃1⁃cysteine，NAC）后，划痕、Transwell小室
观察类胰蛋白酶诱导的细胞修复与迁移功能。结果：类胰蛋白酶呈剂量依赖性地促进16HBE修复与迁移；下调E⁃cadherin、上
调N⁃cadherin及α⁃平滑肌肌动蛋白（α⁃smooth muscle actin，α⁃SMA）表达（P < 0.05）。类胰蛋白酶增加16HBE细胞内ROS水平，
而PAR2抑制剂FSLLRY⁃NH2可逆转上述改变（P < 0.05）。NAC可有效抑制类胰蛋白酶诱导的细胞修复与迁移（P < 0.05）。结
论：激活PAR2可经由ROS信号调控人支气管上皮细胞EMT过程，是细胞表型转化与重塑调控的潜在靶标。
［关键词］ 蛋白酶激活受体2；上皮间充质转化；活性氧；人支气管上皮细胞；类胰蛋白酶
［中图分类号］ R562.25 ［文献标志码］ A ［文章编号］ 1007⁃4368（2022）06⁃790⁃07
doi：10.7655/NYDXBNS20220604

Activation of protease ⁃ activated receptor 2 elicits epithelial ⁃ mesenchymal transition in
human bronchial epithelial cells via ROS signaling

*
WANG Hongyu，GU Hao，GE Ai，ZENG Xiaoning ，YAO Xin *
Department of Respiratory and Critical Care Medicine，the First Affiliated Hospital of Nanjing Medical University，
Nanjing 210029，China

［Abstract］ Objective：This study aims to investigate the role of protease ⁃ activated receptor 2（PAR2）involved in epithelial ⁃
mesenchymal transition（EMT）of human bronchial epithelial（HBE）cells. Methods：The 16HBE cells were stimulated with various
concentrations of tryptase（a nature agonist of PAR2）. Cell migration and repair were assessed by transwell and scratch assay. Western
blot was used to examine the expressions of EMT associated biomarkers including E⁃cadherin，N⁃cadherin and α⁃smooth muscle actin
（α⁃SMA）. DCFH⁃DA probe was employed to measure the generation of reactive oxygen species（ROS）. The effects of N⁃acety⁃1⁃
cysteine（NAC）on 16HBE induced by tryptase were also examined by transwell and scratch assay. Results：Tryptase dramatically
promoted cell migration and repair with loss of E⁃cadherin and increase of N⁃cadherin and α⁃SMA in a dose⁃dependent manner（P <
0.05）. Tryptase enhanced generation of ROS in 16HBE cells，and this effect can be inhibited by PAR2 antagonist FSLLRY⁃NH2（P <
0.05）. ROS scavenger NAC significantly inhibited the EMT changes induced by tryptase（P < 0.05）. Conclusion：Activation of PAR2
triggered EMT process via ROS signal in HBE cells，which highlights a potential target for the regulation of HBE phenotype shift
involved in airway remodeling.
［Key words］ protease⁃activated receptor 2；epithelial⁃mesenchymal transition；ROS；HBE；tryptase
［J Nanjing Med Univ，2022，42（06）：790⁃795，829］

气道重塑是支气管哮喘（简称哮喘）发生发展

［基金项目］ 国家自然科学基金（81970016，81870039） 的重要事件，包括一系列关键环节，如上皮间充质
∗ 转化（epithelial⁃mesenchymal transition，EMT）、成纤
通信作者（Corresponding author），E⁃mail：zeng_xiao_njng@njmu.
edu.cn；yaoxin@njmu.edu.cn 维细胞活化及分化为肌成纤维细胞、细胞外基质