瞬时受体电位香草酸亚型4（TRPV4）通道在视网膜血管疾病中促进血管通透性。

TRPV4 channels promote vascular permeability in retinal vascular disease.

作者信息

Nishinaka Anri, Tanaka Miruto, Ohara Kentaro, Sugaru Eiji, Shishido Yuji, Sugiura Akemi, Moriguchi Yukiko, Toui Amane, Nakamura Shinsuke, Shimada Kaoru, Watanabe Shuzo, Hara Hideaki, Shimazawa Masamitsu

机构信息

Molecular Pharmacology, Department of Biofunctional Evaluation, Gifu Pharmaceutical University, Gifu, Japan.

RaQualia, Pharma Inc., Nagoya, Japan; RaQualia Pharma Industry-Academia Collaborative Research Center, Research Institute of Environmental Medicine, Nagoya University, Nagoya, Japan.

出版信息

Exp Eye Res. 2023 Mar;228:109405. doi: 10.1016/j.exer.2023.109405. Epub 2023 Feb 9.

DOI:10.1016/j.exer.2023.109405

PMID:36773739

Abstract

This study aimed to determine the role of transient receptor potential vanilloid 4 (TRPV4), a calcium (Ca)-permeable cation channel, in the pathophysiology of retinal vascular disease. The retinal vein occlusion (RVO) murine model was created by irradiating retinal veins using lasers. TRPV4 expression and localization were evaluated in RVO mice retinas. In addition, we examined the effects of TRPV4 antagonists (RQ-00317310, HC-067047, GSK2193874, and GSK2798745) on retinal edema, blood flow, and ischemic areas in RVO mice. Furthermore, changes in the retinal expression of tumor necrosis factor (TNF)-α and aquaporin4 (AQP4) by RQ-00317310 were analyzed using Western blot. We also assessed the barrier integrity of epithelial cell monolayers using trans-endothelial electrical resistance (TEER) in Human Retinal Microvascular Endothelial Cells (HRMECs). The expression of TRPV4 was significantly increased and co-localized with glutamine synthetase (GS), a Müller glial marker, in the ganglion cell layer (GCL) of the RVO mice. Moreover, RQ-00317310 administration ameliorated the development of retinal edema and ischemia in RVO mice. In addition, the up regulation of TNF-α and down-regulation of AQP4 were lessened by the treatment with RQ-00317310. Treatment with GSK1016790A, a TRPV4 agonist, increased vascular permeability, while RQ-00317310 treatment decreased vascular endothelial growth factor (VEGF)- or TRPV4-induced retinal vascular hyperpermeability in HRMECs. These findings suggest that TRPV4 plays a role in the development of retinal edema and ischemia. Thus, TRPV4 could be a new therapeutic target against the pathological symptoms of retinal vascular diseases.

摘要

本研究旨在确定瞬时受体电位香草酸受体4（TRPV4），一种钙（Ca）通透性阳离子通道，在视网膜血管疾病病理生理学中的作用。通过激光照射视网膜静脉建立视网膜静脉阻塞（RVO）小鼠模型。评估RVO小鼠视网膜中TRPV4的表达和定位。此外，我们研究了TRPV4拮抗剂（RQ-00317310、HC-067047、GSK2193874和GSK2798745）对RVO小鼠视网膜水肿、血流和缺血区域的影响。此外，使用蛋白质免疫印迹法分析RQ-00317310对视网膜肿瘤坏死因子（TNF）-α和水通道蛋白4（AQP4）表达的影响。我们还使用人视网膜微血管内皮细胞（HRMECs）的跨内皮电阻（TEER）评估上皮细胞单层的屏障完整性。在RVO小鼠的神经节细胞层（GCL）中，TRPV4的表达显著增加，并与穆勒胶质细胞标志物谷氨酰胺合成酶（GS）共定位。此外，给予RQ-00317310可改善RVO小鼠视网膜水肿和缺血的发展。此外，RQ-00317310治疗可减轻TNF-α的上调和AQP4的下调。用TRPV4激动剂GSK1016790A处理可增加血管通透性，而RQ-00317310处理可降低血管内皮生长因子（VEGF）或TRPV4诱导的HRMECs视网膜血管高通透性。这些发现表明TRPV4在视网膜水肿和缺血的发展中起作用。因此，TRPV4可能是针对视网膜血管疾病病理症状的新治疗靶点。