Wen Lei, Wen Yue-Chun, Ke Gen-Jie, Sun Si-Qin, Dong Kai, Wang Lin, Liao Rong-Feng
Department of Ophthalmology, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China.
Department of Ophthalmology, Anhui Provincial Hospital, Hefei, Anhui, China.
BMC Ophthalmol. 2018 Feb 12;18(1):38. doi: 10.1186/s12886-018-0697-2.
Ca entry plays an important role in modulating endothelial cell migration and tube formation. Transient receptor potential cation channel subfamily V member 4 (TRPV4) is a Ca-permeable channel that is widely expressed in endothelial cells. It has been reported that TRPV4 is expressed in HRCECs and regulates Ca entry. However, the function of TRPV4 in human retinal capillary endothelial cells (HRCECs) remains unknown.
In this study we used western blot and immunostaining assay to verify TRPV4 expression in HRCECs. And then we pretreated HRCECs with HC067047 and transfected with specific shRNA of TRPV4. The functional presence of TrpV4 was determined by using fluorescence, migration and tube formation assay in TrpV4 knockdown cells or control cells.
Using western blot and immunostaining, we confirmed TRPV4 expression in HRCECs. Moreover, inhibition of TRPV4 using the specific inhibitor HC067047 and the knockdown of TRPV4 with shRNA significantly suppressed tube formation and migration by HRCECs.
TRPV4 is essential for HRCEC migration and tube formation, and maybe a potential therapeutic target for retinal vascular diseases.
钙内流在调节内皮细胞迁移和管腔形成中起重要作用。瞬时受体电位阳离子通道亚家族V成员4（TRPV4）是一种钙通透通道，在内皮细胞中广泛表达。据报道，TRPV4在人视网膜毛细血管内皮细胞（HRCECs）中表达并调节钙内流。然而，TRPV4在人视网膜毛细血管内皮细胞中的功能仍不清楚。
在本研究中，我们使用蛋白质印迹法和免疫染色法验证TRPV4在HRCECs中的表达。然后我们用HC067047预处理HRCECs并用TRPV4特异性短发夹RNA进行转染。通过在TRPV4敲低细胞或对照细胞中使用荧光、迁移和管腔形成试验来确定TrpV4的功能存在。
使用蛋白质印迹法和免疫染色法，我们证实了TRPV4在HRCECs中的表达。此外，使用特异性抑制剂HC067047抑制TRPV4以及用短发夹RNA敲低TRPV4可显著抑制HRCECs的管腔形成和迁移。
TRPV4对HRCEC迁移和管腔形成至关重要，可能是视网膜血管疾病的潜在治疗靶点。
