Dunne Orla M, Martin S Lorraine, Sergeant Gerard P, McAuley Daniel F, O'Kane Cecilia M, Button Brian, McGarvey Lorcan P, Lundy Fionnuala T
Wellcome-Wolfson Institute for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, 97 Lisburn Road, Belfast, BT9 7BL, UK.
School of Pharmacy, Queen's University Belfast, Belfast, UK.
Respir Res. 2024 Apr 27;25(1):188. doi: 10.1186/s12931-024-02807-0.
Repetitive bouts of coughing expose the large airways to significant cycles of shear stress. This leads to the release of alarmins and the tussive agent adenosine triphosphate (ATP) which may be modulated by the activity of ion channels present in the human airway. This study aimed to investigate the role of the transient receptor potential subfamily vanilloid member 2 (TRPV2) channel in mechanically induced ATP release from primary bronchial epithelial cells (PBECs).PBECs were obtained from individuals undergoing bronchoscopy. They were cultured in vitro and exposed to mechanical stress in the form of compressive and fluid shear stress (CFSS) or fluid shear stress (FSS) alone at various intensities. ATP release was measured using a luciferin-luciferase assay. Functional TRPV2 protein expression in human PBECs was investigated by confocal calcium imaging. The role of TRPV2 inhibition on FSS-induced ATP release was investigated using the TRPV2 inhibitor tranilast or siRNA knockdown of TRPV2. TRPV2 protein expression in human lung tissue was also determined by immunohistochemistry.ATP release was significantly increased in PBECs subjected to CFSS compared with control (unstimulated) PBECs (N = 3, ***P < 0.001). PBECs expressed functional TRPV2 channels. TRPV2 protein was also detected in fixed human lung tissue. ATP release from FFS stimulated PBECs was decreased by the TRPV2 inhibitor tranilast (N = 3, **P < 0.01) (vehicle: 159 ± 17.49 nM, tranilast: 25.08 ± 5.1 nM) or by TRPV2 siRNA knockdown (N = 3, *P < 0.05) (vehicle: 197 ± 24.52 nM, siRNA: 119 ± 26.85 nM).In conclusion, TRPV2 is expressed in the human airway and modulates ATP release from mechanically stimulated PBECs.
反复咳嗽会使大气道暴露于显著的剪切应力循环中。这会导致警报素和咳嗽介质三磷酸腺苷（ATP）的释放，而这可能会受到人类气道中存在的离子通道活性的调节。本研究旨在探讨瞬时受体电位香草酸亚家族成员2（TRPV2）通道在机械诱导的原代支气管上皮细胞（PBECs）ATP释放中的作用。
PBECs取自接受支气管镜检查的个体。将它们在体外培养，并以压缩和流体剪切应力（CFSS）或单独的流体剪切应力（FSS）形式，在不同强度下暴露于机械应力。使用荧光素 - 荧光素酶测定法测量ATP释放。通过共聚焦钙成像研究人PBECs中功能性TRPV2蛋白的表达。使用TRPV2抑制剂曲尼司特或TRPV2的siRNA敲低来研究TRPV2抑制对FSS诱导的ATP释放的作用。还通过免疫组织化学测定人肺组织中TRPV2蛋白的表达。
与对照（未刺激）PBECs相比，经受CFSS的PBECs中ATP释放显著增加（N = 3，***P < 0.001）。PBECs表达功能性TRPV2通道。在固定的人肺组织中也检测到TRPV2蛋白。TRPV2抑制剂曲尼司特（N = 3，**P < 0.01）（载体：159 ± 17.49 nM，曲尼司特：25.08 ± 5.1 nM）或TRPV2 siRNA敲低（N = 3，*P < 0.05）（载体：197 ± 24.52 nM，siRNA：119 ± 26.85 nM）可降低FSS刺激的PBECs中的ATP释放。
总之，TRPV2在人类气道中表达，并调节机械刺激的PBECs中的ATP释放。
